TOP TEN perturbations for 1553088_a_at (Homo sapiens)

Organism: Homo sapiens
Gene: 1553088_a_at
Selected probe(set): 225606_at
Platform: Affymetrix Human Genome U133 Plus 2.0 Array

Expression of 1553088_a_at (225606_at) across 6673 perturbations tested by GENEVESTIGATOR:

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

Relative Expression (log2-ratio):4.7153206
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 (4h) / control virus transfected EREB2-5 cell sample (24h)

Relative Expression (log2-ratio):4.2779875
Number of Samples:3 / 3
Experimental 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.
Control control virus transfected EREB2-5 cell sample (24h)
EREB2-5 cell line stably transfected with control expression plasmid coding for bacterial chloramphenicol acetyltransferase (CAT) gene. CAT was used as a negative control. Briefly, before the CAT induction, cells were maintained in RPMI media and deprived of estrogen for 3 days before doxycycline was added. 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. Estrogen withdrawal leads to the inactivation of EBNA2, resulting in cell cycle arrest. Before the preparation of RNA, NGFR cells were purified by MACS separation to enrich the cells with a transcriptional response to doxycycline.

CD44s overexpr. study 1 / normal HEK-293 cell sample

Relative Expression (log2-ratio):-4.209408
Number of Samples:3 / 3
Experimental CD44s overexpr. study 1
Human embryonic kidney cell line HEK-293 transfected with pcDNA3.1(-) vector containing codon-optimized human CD44 sequence for expression.
Control normal HEK-293 cell sample
Untransfected, native human embryonic kidney cell line HEK-293 cell samples.

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

Relative Expression (log2-ratio):4.19934
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 (re-differentiated; PPRF; 4d) / pancreatic islet study 3 (expanded; PPRF)

Relative Expression (log2-ratio):4.1943855
Number of Samples:2 / 2
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 4d)
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 4 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 1.0ug/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):-4.01087
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.

pancreatic islet study 3 (re-differentiated; PPRF; 2d) / pancreatic islet study 3 (expanded; PPRF)

Relative Expression (log2-ratio):3.9561205
Number of Samples:2 / 2
Experimental pancreatic islet study 3 (re-differentiated; PPRF; 2d)
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 2 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 1.0ug/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.

dendritic cell study 6 (gardiquimod) / dendritic cell study 6 (untreated)

Relative Expression (log2-ratio):3.7026396
Number of Samples:7 / 8
Experimental dendritic cell study 6 (gardiquimod)
Plasmacytoid dendritic cells (pDCs) collected from blood of healthy donors and treated with gardiquimod (TLR7 agonist) for 3 hours. pDCs were sorted as CD303+, CD123+, CD45+, CD69+, CD40+, CD86+.
Control dendritic cell study 6 (untreated)
Plasmacytoid dendritic cells (pDCs) collected from blood of healthy donors. pDCs were sorted as CD303+, CD123+, CD45+, CD69+, CD40+, CD86+.

dendritic cell study 6 (gardiquimod; RN486) / dendritic cell study 6 (CpG A; RN486)

Relative Expression (log2-ratio):3.6936922
Number of Samples:4 / 4
Experimental dendritic cell study 6 (gardiquimod; RN486)
Plasmacytoid dendritic cells (pDCs) collected from blood of healthy donors and treated with gardiquimod (TLR7 agonist) and RN486 (Bruton’s tyrosine kinase; BTK inhibitor) for 3 hours. pDCs were sorted as CD303+, CD123+, CD45+, CD69+, CD40+, CD86+.
Control dendritic cell study 6 (CpG A; RN486)
Plasmacytoid dendritic cells (pDCs) collected from blood of healthy donors and treated with CpG-A ODN2216 Class A (TLR9 agonist) and RN486 (Bruton’s tyrosine kinase; BTK inhibitor) for 3 hours. pDCs were sorted as CD303+, CD123+, CD45+, CD69+, CD40+, CD86+.

dendritic cell study 6 (gardiquimod; RN486) / dendritic cell study 6 (untreated)

Relative Expression (log2-ratio):3.6601992
Number of Samples:4 / 8
Experimental dendritic cell study 6 (gardiquimod; RN486)
Plasmacytoid dendritic cells (pDCs) collected from blood of healthy donors and treated with gardiquimod (TLR7 agonist) and RN486 (Bruton’s tyrosine kinase; BTK inhibitor) for 3 hours. pDCs were sorted as CD303+, CD123+, CD45+, CD69+, CD40+, CD86+.
Control dendritic cell study 6 (untreated)
Plasmacytoid dendritic cells (pDCs) collected from blood of healthy donors. pDCs were sorted as CD303+, CD123+, CD45+, CD69+, CD40+, CD86+.