Journal: Physiological Genomics

Article Title: Gene expression effects of glucocorticoid and mineralocorticoid receptor agonists and antagonists on normal human skeletal muscle

doi: 10.1152/physiolgenomics.00128.2016

Figure Lengend Snippet: Treatment with eplerenone or mifepristone alone caused gene expression changes in normal human myotubes. A: treatment with the selective MR antagonist eplerenone resulted in 42 gene expression changes and functional clustering of these genes revealed: 3 apoptotic, 5 immune or defense response, 3 transcriptional regulators, 4 ion binding, 3 cell adhesion, 3 alternative splicing, 12 muscle contraction, and 9 genes with unknown or specific functions. B: treatment with the GR antagonist mifepristone resulted in 27 gene expression changes and functional clustering of these genes revealed: 1 apoptotic, 4 immune or defense response, 9 regulators of transcription, 2 ion binding, 2 alternative splicing, 2 developmental, and 7 genes with unknown or specific functions.

Article Snippet: Cells were serum restricted for 5 days in high-glucose Dulbecco’s modified medium (DMEM, Invitrogen) and supplemented with 2% horse serum and 100 U/ml penicillin-streptomycin, followed by 48 h treatments with: aldosterone (10 µM; MR EC 50 : 1.3 nM; GR EC 50 : 80 nM), eplerenone (10 µM; MR IC 50 : 81 nM; Pfizer Compound Transfer Program), mifepristone (1 µM; GR IC 50 : 2.6 nM; Cayman Chemical), prednisolone (1 µM; GR EC 50 : 4.4 nM), spironolactone (10 µM; MR IC 50 : 1.6 nM; GR IC 50 : 2.9 µM), or vehicle (drugs were purchased from Sigma and dissolved in 100% DMSO unless specified otherwise).

Techniques: Expressing, Functional Assay, Binding Assay

Journal: Physiological Genomics

Article Title: Gene expression effects of glucocorticoid and mineralocorticoid receptor agonists and antagonists on normal human skeletal muscle

doi: 10.1152/physiolgenomics.00128.2016

Figure Lengend Snippet: Gene expression changes with low-dose aldosterone compared with aldosterone plus antagonists in normal human myotubes. A: co-incubation with aldosterone plus eplerenone increased 8 genes and decreased 85 genes (93 total) compared with myotubes treated with aldosterone alone. Functional clustering revealed: 4 apoptotic, 7 transcriptional regulators, 5 ion binding, 7 transmembrane, 6 cell adhesion, 11 cytoskeletal protein binding, 4 muscle contraction, 27 cell cycle, and 22 genes with unknown or specific functions. B: co-incubation with aldosterone plus spironolactone increased 24 genes and decreased 90 genes (114 total) compared with myotubes treated with aldosterone alone. Functional clustering revealed: 9 apoptotic, 8 immune or defense response, 4 transcriptional regulators, 9 ion binding, 6 transmembrane, 13 cell adhesion, 8 structural constituents of muscle, 30 tissue development, and 27 genes with unknown or specific functions. C: co-incubation with aldosterone plus mifepristone increased 251 genes and decreased 370 genes (621 total) compared with myotubes treated with aldosterone alone. Functional clustering revealed: 33 apoptotic, 38 immune or defense response, 62 transcriptional regulators, 63 ion binding, 45 transmembrane, 18 cell adhesion, 63 ECM or cytoskeletal protein binding, 30 alternative splicing, 106 vasculature or muscle structure development, 25 oxidative stress responsive, 54 cell cycle, 7 oxidoreductase, 9 GTPase, and 68 genes with unknown or specific functions.

Article Snippet: Cells were serum restricted for 5 days in high-glucose Dulbecco’s modified medium (DMEM, Invitrogen) and supplemented with 2% horse serum and 100 U/ml penicillin-streptomycin, followed by 48 h treatments with: aldosterone (10 µM; MR EC 50 : 1.3 nM; GR EC 50 : 80 nM), eplerenone (10 µM; MR IC 50 : 81 nM; Pfizer Compound Transfer Program), mifepristone (1 µM; GR IC 50 : 2.6 nM; Cayman Chemical), prednisolone (1 µM; GR EC 50 : 4.4 nM), spironolactone (10 µM; MR IC 50 : 1.6 nM; GR IC 50 : 2.9 µM), or vehicle (drugs were purchased from Sigma and dissolved in 100% DMSO unless specified otherwise).

Techniques: Expressing, Incubation, Functional Assay, Binding Assay, Protein Binding