TOP TEN perturbations for P52732 (Homo sapiens)

Organism: Homo sapiens
Gene: P52732
Selected probe(set): 204444_at
Platform: Affymetrix Human Genome U133 Plus 2.0 Array

Expression of P52732 (204444_at) across 5217 perturbations tested by GENEVESTIGATOR:

pancreatic islet study 3 (re-differentiated; PPRF; 8d) / pancreatic islet study 3 (expanded; PPRF)

Relative Expression (log2-ratio):-5.4189453
Number of Samples:2 / 2
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 8d)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded for 6 weeks and re-differentiated for 8 days according to Pharmaceutical Production Research Facility (PPRF) Protocol. Re-differentiation phase: For induction of expanded cells to differentiate into islet-like cell clusters, the cells were seeded into 60 mm ultra-low attachment dishes in serum-free CMRL-1066 supplemented with 4 mM L-glutamine, 1% BSA, insulin (10 g/ml), transferrin (5.5 g/ml), sodium selenite (6.7ng/ml), and 1% antibiotic antimycotic solution at a density of 6.0 x 10EXP6 cells per dish. The induction medium was further added with islet neogenesis-associated protein (INGAP) peptide at a concentration of 1.0 ug/ml.
Control pancreatic islet study 3 (expanded; PPRF)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded according to Pharmaceutical Production Research Facility Protocol (PPRF) for 6 weeks. Expansion phase: Approximately 2,500 islet equivalents were seeded onto 75-cm2 fibronectin-coated tissue culture-treated flasks in a proprietary serum-free medium (PPRF medium). The serum-free PPRF medium was supplemented with 30% serum-free mesenchymal-conditioned medium. When the cell migration resulted in the formation of large-size colonies, the cells were harvested by trypsinization and replated into new tissue culture flasks. Once the cells reached near confluence (80?90%) in monolayer, they were trypsinized, counted, and subcultured at a density of 5,000 cells per cm2. Thereafter, cells were serially passaged using the same protocol.

pancreatic islet study 3 (expanded; PPRF) / normal pancreatic islet sample

Relative Expression (log2-ratio):5.241623
Number of Samples:2 / 7
Experimental pancreatic islet study 3 (expanded; PPRF)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded according to Pharmaceutical Production Research Facility Protocol (PPRF) for 6 weeks. Expansion phase: Approximately 2,500 islet equivalents were seeded onto 75-cm2 fibronectin-coated tissue culture-treated flasks in a proprietary serum-free medium (PPRF medium). The serum-free PPRF medium was supplemented with 30% serum-free mesenchymal-conditioned medium. When the cell migration resulted in the formation of large-size colonies, the cells were harvested by trypsinization and replated into new tissue culture flasks. Once the cells reached near confluence (80?90%) in monolayer, they were trypsinized, counted, and subcultured at a density of 5,000 cells per cm2. Thereafter, cells were serially passaged using the same protocol.
Control normal pancreatic islet sample
Pancreatic islets were obtained from seven donors aged between 37 and 70 years and body mass index between 22 and 27. Functional islets were cultured in CMRL 1066 supplemented with 10% FCS, 1% glutamine, 5.6 mM glucose, 1 mM HEPES, 110 U/ml penicillin, and 110 g/ml streptomycin for 7 days or immediately processed after isolation.

nutlin-3 (10uM); flagellin (500 ng/ml) study 1 (control vector) / flagellin study 4 (control vector; 500 ng/ml)

Relative Expression (log2-ratio):-5.002784
Number of Samples:6 / 2
Experimental nutlin-3 (10uM); flagellin (500 ng/ml) study 1 (control vector)
Human breast adenocarcinoma cells MCF-7 expressing wild type p53, stably transfected with control empty vector pSUPER, treated with nutlin-3 (10 uM) for 45 hours followed by an additional 3 hours treatment of nutlin-3 (10uM) and flagellin (500 ng/ml). Cells were cultured in RPMI-1640 medium supplemented with 10% heat inactivated FBS and 50 U/ml of penicillin, 50 ug/ml of streptomycin. ATC code:---
Control flagellin study 4 (control vector; 500 ng/ml)
Human breast adenocarcinoma cells MCF-7 expressing wild type p53, stably transfected with control empty vector pSUPER, treated with DMSO 0.1% for 45 hours followed by an additional 3 hours with flagellin (500 ng/ml). Cells were cultured in RPMI-1640 medium supplemented with 10% heat inactivated FBS and 50 U/ml of penicillin, 50 ug/ml of streptomycin.

stem cell differentiation study 47 (BMP-2; TGFb; 2d) / stem cell differentiation study 47 (0d)

Relative Expression (log2-ratio):-4.9702425
Number of Samples:3 / 6
Experimental stem cell differentiation study 47 (BMP-2; TGFb; 2d)
Bone marrow-derived mesenchymal stem cell line (MSC) differentiated for 2 days in the presence of bone morphogenetic protein 2 (BMP-2, 50ng/ml) and transforming growth factor beta (TGFb, 5 ng/ml). Cells were propagated for not more than five passages in mesenchymal stem cell growth medium, at 37 ?C and in a humidified atmosphere containing 7.5 % CO2. MSCs were incubated for 24 hours in proliferation medium (PM, high glucose DMEM, 10 % FBS, 100 U/ml penicillin, and 100 ?g/ml streptomycin). Subsequently PMCs were differentiated for 2 days in differentiation medium (consisting of PM with 10EXP(-6)M dexamethasone, 10 ?g/ml insulin, 10EXP(-7) M rosiglitazone, 50 ng/ml BMP-2, 5 ng/ml TGFb), and harvested. Samples are biological replicates. Normal bone marrow was obtained from 3 healthy donors (5F0138, 6F4085, 7F3458).
Control stem cell differentiation study 47 (0d)
Undifferentiated bone marrow-derived mesenchymal stem cell line (MSC) without any treatment. Cells were propagated for not more than five passages in mesenchymal stem cell growth medium, at 37 ?C and in a humidified atmosphere containing 7.5 % CO2. MSCs were incubated for 24 hours in proliferation medium (high glucose DMEM, 10 % FBS, 100 U/ml penicillin, and 100 ?g/ml streptomycin) and harvested. Samples are biological replicates. Normal bone marrow was obtained from 3 healthy donors (5F0138, 6F4085, 7F3458).

stem cell differentiation study 47 (BMP-2; TGFb; 3d) / stem cell differentiation study 47 (0d)

Relative Expression (log2-ratio):-4.8537884
Number of Samples:3 / 6
Experimental stem cell differentiation study 47 (BMP-2; TGFb; 3d)
Bone marrow-derived mesenchymal stem cell line (MSC) differentiated for 3 days in the presence of bone morphogenetic protein 2 (BMP-2, 50ng/ml) and transforming growth factor beta (TGFb, 5 ng/ml). Cells were propagated for not more than five passages in mesenchymal stem cell growth medium, at 37 ?C and in a humidified atmosphere containing 7.5 % CO2. MSCs were incubated for 24 hours in proliferation medium (PM, high glucose DMEM, 10 % FBS, 100 U/ml penicillin, and 100 ?g/ml streptomycin). Subsequently PMCs were differentiated for 3 days in differentiation medium (consisting of PM with 10EXP(-6)M dexamethasone, 10 ?g/ml insulin, 10EXP(-7) M rosiglitazone, 50 ng/ml BMP-2, 5 ng/ml TGFb), and harvested. Samples are biological replicates. Normal bone marrow was obtained from 3 healthy donors (5F0138, 6F4085, 7F3458).
Control stem cell differentiation study 47 (0d)
Undifferentiated bone marrow-derived mesenchymal stem cell line (MSC) without any treatment. Cells were propagated for not more than five passages in mesenchymal stem cell growth medium, at 37 ?C and in a humidified atmosphere containing 7.5 % CO2. MSCs were incubated for 24 hours in proliferation medium (high glucose DMEM, 10 % FBS, 100 U/ml penicillin, and 100 ?g/ml streptomycin) and harvested. Samples are biological replicates. Normal bone marrow was obtained from 3 healthy donors (5F0138, 6F4085, 7F3458).

pancreatic islet study 3 (re-differentiated; PPRF; 6d) / pancreatic islet study 3 (expanded; PPRF)

Relative Expression (log2-ratio):-4.7317247
Number of Samples:2 / 2
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 6d)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded for 6 weeks and re-differentiated for 6 days according to Pharmaceutical Production Research Facility (PPRF) Protocol. Re-differentiation phase: For induction of expanded cells to differentiate into islet-like cell clusters, the cells were seeded into 60 mm ultra-low attachment dishes in serum-free CMRL-1066 supplemented with 4 mM L-glutamine, 1% BSA, insulin (10 g/ml), transferrin (5.5 g/ml), sodium selenite (6.7ng/ml), and 1% antibiotic antimycotic solution at a density of 6.0 x 10EXP6 cells per dish. The induction medium was further added with islet neogenesis-associated protein (INGAP) peptide at a concentration of 1.0 ug/ml.
Control pancreatic islet study 3 (expanded; PPRF)
Human pancreatic islets were isolated from a cadaveric donor. The population was 70% pure in mature islets, which was determined by dithizone staining. Islets cells were expanded according to Pharmaceutical Production Research Facility Protocol (PPRF) for 6 weeks. Expansion phase: Approximately 2,500 islet equivalents were seeded onto 75-cm2 fibronectin-coated tissue culture-treated flasks in a proprietary serum-free medium (PPRF medium). The serum-free PPRF medium was supplemented with 30% serum-free mesenchymal-conditioned medium. When the cell migration resulted in the formation of large-size colonies, the cells were harvested by trypsinization and replated into new tissue culture flasks. Once the cells reached near confluence (80?90%) in monolayer, they were trypsinized, counted, and subcultured at a density of 5,000 cells per cm2. Thereafter, cells were serially passaged using the same protocol.

EBNA2 overexpr. study 1 (24h) / EBNA2 overexpr. study 1 (4h)

Relative Expression (log2-ratio):4.7277584
Number of Samples:3 / 3
Experimental EBNA2 overexpr. study 1 (24h)
EREB2-5 cell line stably transfected with the chimeric Epstein-Barr virus-encoded nuclear antigen 2 (EBNA2) and control expression plasmid coding for bacterial chloramphenicol acetyltransferase (CAT) gene and . CAT was used as a negative control. Briefly, before the CAT and EBNA2 induction, cells were maintained in RPMI media and deprived of estrogen for 3 days before doxycycline and estrogen was added. Expression of EBNA2 was induced after addition of estrogen and expression of CAT was induced by the addition of doxycycline to a final concentration of 100 ng/mL medium. Doxycycline induced cells were harvested 24 hours after induction. Because EBNA2 is essential for the proliferation of EBV-infected cells, EREB2-5 cells grow only in the presence of estrogen. Before the preparation of RNA, NGFR cells were purified by MACS separation to enrich the cells with a transcriptional response to doxycycline.
Control EBNA2 overexpr. study 1 (4h)
EREB2-5 cell line stably transfected with the chimeric Epstein-Barr virus-encoded nuclear antigen 2 (EBNA2) and control expression plasmid coding for bacterial chloramphenicol acetyltransferase (CAT) gene and . CAT was used as a negative control. Briefly, before the CAT and EBNA2 induction, cells were maintained in RPMI media and deprived of estrogen for 3 days before doxycycline and estrogen was added. Expression of EBNA2 was induced after addition of estrogen and expression of CAT was induced by the addition of doxycycline to a final concentration of 100 ng/mL medium. Doxycycline induced cells were harvested 4 hours after induction. Because EBNA2 is essential for the proliferation of EBV-infected cells, EREB2-5 cells grow only in the presence of estrogen. Before the preparation of RNA, NGFR cells were purified by MACS separation to enrich the cells with a transcriptional response to doxycycline.

stem cell differentiation study 47 (BMP-2; IBMX; 7d) / stem cell differentiation study 47 (0d)

Relative Expression (log2-ratio):-4.643261
Number of Samples:3 / 6
Experimental stem cell differentiation study 47 (BMP-2; IBMX; 7d)
Bone marrow-derived mesenchymal stem cell line (MSC) differentiated for 7 days in the presence of bone morphogenetic protein 2 (BMP-2, 50ng/ml) and 3-isobutyl-1-methylxanthine (IBMX, 250 ?M). Cells were propagated for not more than five passages in mesenchymal stem cell growth medium, at 37 ?C and in a humidified atmosphere containing 7.5 % CO2. MSCs were incubated for 24 hours in proliferation medium (PM, high glucose DMEM, 10 % FBS, 100 U/ml penicillin, and 100 ?g/ml streptomycin). Subsequently PMCs were differentiated for 7 days in differentiation medium (consisting of PM with 10EXP(-6)M dexamethasone, 10 ?g/ml insulin, 10EXP(-7) M rosiglitazone, 50 ng/ml BMP-2, 250 ?M IBMX), and harvested. Samples are biological replicates. Normal bone marrow was obtained from 3 healthy donors (5F0138, 6F4085, 7F3458).
Control stem cell differentiation study 47 (0d)
Undifferentiated bone marrow-derived mesenchymal stem cell line (MSC) without any treatment. Cells were propagated for not more than five passages in mesenchymal stem cell growth medium, at 37 ?C and in a humidified atmosphere containing 7.5 % CO2. MSCs were incubated for 24 hours in proliferation medium (high glucose DMEM, 10 % FBS, 100 U/ml penicillin, and 100 ?g/ml streptomycin) and harvested. Samples are biological replicates. Normal bone marrow was obtained from 3 healthy donors (5F0138, 6F4085, 7F3458).

nutlin-3 study 2 (control vector; 10uM) / vehicle treated MCF-7-con cell sample

Relative Expression (log2-ratio):-4.5881414
Number of Samples:6 / 3
Experimental nutlin-3 study 2 (control vector; 10uM)
Human breast adenocarcinoma cells MCF-7 expressing wild type p53, stably transfected with control empty vector pSUPER, treated with nutlin-3 (10 uM) for 48 hours. Cells were cultured in RPMI-1640 medium supplemented with 10% heat inactivated FBS and 50 U/ml of penicillin, 50 ug/ml of streptomycin. ATC code:---
Control vehicle treated MCF-7-con cell sample
Human breast adenocarcinoma cells MCF-7 expressing wild type p53 stably transfected with control empty vector pSUPER, vehicle treated with DMSO (0.1%) for 48 hours. Cells were cultured in RPMI-1640 medium supplemented with 10% heat inactivated FBS and 50 U/ml of penicillin, 50 ug/ml of streptomycin.

keratinocyte differentiation study 2 (KLF4 siRNA) / keratinocyte differentiation study 2

Relative Expression (log2-ratio):4.533059
Number of Samples:2 / 2
Experimental keratinocyte differentiation study 2 (KLF4 siRNA)
KLF4 (Kruppel-like factor 4) depleted primary neonatal keratinocytes differentiated with 1.2 mM calcium for 3 days. Keratinocytes obtained from freshly isolated foreskin were grown in keratinocyte-SFM medium supplemented with epidermal growth factor and bovine pituitary extract. KLF4 depletion was done using siRNAs: KLF4i(A): CCGAGGAGTTCAACGATCT; KLF4i(B): TGACCAGGCACTACCGTAA. Differentiation was induced at 100% confluency.
Control keratinocyte differentiation study 2
Primary neonatal keratinocytes differentiated with 1.2 mM calcium for 3 days. Keratinocytes obtained from freshly isolated foreskin were grown in keratinocyte-SFM medium supplemented with epidermal growth factor and bovine pituitary extract. Differentiation was induced at 100% confluency.