Journal: Hormones & Cancer

Article Title: CREB-Regulated Transcription Co-Activator Family Stimulates Promoter II-Driven Aromatase Expression in Preadipocytes

doi: 10.1007/s12672-013-0142-1

Figure Lengend Snippet: Role of CRTCs in aromatase PII activation. a Relative abundance of CRTC transcripts in primary breast preadipocytes with and without PGE2 treatment. b Confocal images of immunofluorescence on endogenous CRTC proteins (green) in primary breast preadipocytes with and without PGE2 treatment, lamin B1 + B2 nuclear stain in red. c Chromatin immunoprecipitation (ChIP) showing endogenous binding of CRTCs to aromatase PII in human breast preadipocytes with and without FSK/PMA treatment. d Reporter assays demonstrating the effect of CRTCs on aromatase PII activity with and without FSK/PMA treatment. vc Vehicle control; mean ± SEM, n = 3 for qPCR and reporter assays; confocal images are representative of the majority of cells examined; scale bar 20.0 μm; all experiments repeated twice

Article Snippet: Cell culture Primary human breast preadipocytes were isolated after collagenase treatment of whole subcutaneous adipose tissue obtained from women undergoing reduction mammoplasty and cultured in Waymouth’s medium (Invitrogen, USA), as previously described [ 19 ].

Techniques: Activation Assay, Immunofluorescence, Staining, Chromatin Immunoprecipitation, Binding Assay, Activity Assay, Control

Journal: Hormones & Cancer

Article Title: CREB-Regulated Transcription Co-Activator Family Stimulates Promoter II-Driven Aromatase Expression in Preadipocytes

doi: 10.1007/s12672-013-0142-1

Figure Lengend Snippet: Role of CRTCs in regulating aromatase expression and activity in primary human breast preadipocytes. Aromatase activity assays demonstrating the effect of silencing (a) and overexpressing (b) CRTCs on the FSK/PMA-mediated stimulation of aromatase activity. c Effect of CRTC overexpression on aromatase transcript expression. Mean ± SEM, n = 3, repeated twice

Article Snippet: Cell culture Primary human breast preadipocytes were isolated after collagenase treatment of whole subcutaneous adipose tissue obtained from women undergoing reduction mammoplasty and cultured in Waymouth’s medium (Invitrogen, USA), as previously described [ 19 ].

Techniques: Expressing, Activity Assay, Over Expression

Journal: Hormones & Cancer

Article Title: CREB-Regulated Transcription Co-Activator Family Stimulates Promoter II-Driven Aromatase Expression in Preadipocytes

doi: 10.1007/s12672-013-0142-1

Figure Lengend Snippet: Effect of phosphorylating CRTC2 at Ser171 on its subcellular localization and the activation of aromatase PII in preadipocytes. a Confocal images on overexpressed GFP-tagged wt, S171A and S171D of CRTC2 (green) and immunofluorescence on lamin B1 + B2 nuclear stain in red, with or without FSK/PMA or AICAR treatment. b Reporter assays demonstrating the effect of wt, S171A and S171D of CRTC2 on aromatase PII activity with or without FSK/PMA treatment. vc Vehicle control, RLU relative luciferase units, β-gal β-galactosidase activity. Mean ± SEM, n = 3 for reporter assays, confocal images are representative of the majority of cells examined, scale bar represents 50.0 μm, all experiments repeated twice

Article Snippet: Cell culture Primary human breast preadipocytes were isolated after collagenase treatment of whole subcutaneous adipose tissue obtained from women undergoing reduction mammoplasty and cultured in Waymouth’s medium (Invitrogen, USA), as previously described [ 19 ].

Techniques: Activation Assay, Immunofluorescence, Staining, Activity Assay, Control, Luciferase