Fig. 2

The C-terminal domain CA of JMY inhibits its ability of MRTF-SRF activation. The mouse fibroblast cell line NIH 3 T3 was transfected with the indicated JMY constructs and analyzed for MRTF-SRF activation under serum-starved conditions. a: Expression of myc-tagged JMY variants was controlled by immunoblotting with myc-specific antibody and tubulin. b: Relative MRTF-SRF luciferase reporter activity upon overexpression of myc-tagged JMY constructs. c: Nuclear translocation of endogenous MRTF-A in myc-JMY-overexpressing cells. Following serum starvation, cells were immuno-stained for endogenous MRTF-A localization. Arrows indicate myc-positive cells. Scale bars, 20 μm. d: Quantification of C by counting 50 myc-expressing cells for each of three independent experiments. e: MRTF-SRF-mediated transcription of smooth muscle α-actin (Acta2) in JMY-expressing cells. f: Organization of the actin cytoskeleton in myc-JMY-overexpressing cells. Following serum-starvation, cells were immunostained for endogenous actin. Arrows indicate myc-positive cells. Scale bars, 20 μm. All data were normalized to the serum-starved control which is set to 1 in b and e. Error bars, s.e.m., n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 according to an unpaired two sample student’s t-test (d) or an unpaired one sample student’s t-test (b, e))