Fig. 2

Myoglobin propagates YAP phosphorylation and inhibits its activity (A) Representative western Blot and quantification of phospho-YAP (pYAP) and total YAP (T-YAP) in control and Mb− cells 48 h after knockdown, quantified as fold change of control; N = 4. (B) Representative western blot and quantitation of pYAP and T-YAP levels in cells with Mb knockdown (control), followed by overexpression of Mb or Apo-Mb for 48 h using GFP tagged Mb or Apo-Mb plasmid. N = 4. (C) Representative immunofluorescence images depicting YAP cellular localization in control and Mb− H9C2 cells. Arrows point to instance of cytoplasmic signal (Ctrl) and nuclear signal (Mb− image). Quantification is the ratio of nuclear to cytoplasmic staining intensity of YAP from 6 high-power field per sample; N = 3. Scale bar: 100 μm. (D) TEAD-luciferase reporter signal in Mb− expressed as fold change of control cells; N = 3. (E) Fold change over control in relative mRNA expression of YAP transcriptional target genes -ctgf and cyr61-in control and Mb− cells; N = 3. Data are mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, n.s. = not significant.