Figure 4.

MRTF-B increases CHRM3 immunoreactive bands in human coronary artery smooth muscle cells. Protein lysates from human coronary artery SMCs treated with null virus and with MRTF-B virus for 120h, respectively, were used for Western blotting using an antibody raised against a human CHRM3 in (A). After developing the blot, it was stripped and incubated with HSP90 antibody (shown below) to assess equal protein loading. At least five bands were detected with the CHRM3 antibody, and three bands at ≈170, ≈140, and ≈45kDa changed significantly with MRTF-B as shown in the compiled analysis in (B). Quantification in panel (B) was done using the blots in (A). Proteins from the same original lysates were also prepared in non-reducing conditions, to examine if the relationship between bands changed (C). The volume was insufficient for one of the null samples in this experiment (lane 6). Non-reducing conditions favored the 140kDa band at the expense of the 45kDa band (D), suggesting multimerization. ****p<0.0001, ***p<0.001, and *p<0.05.