Figure 2.

Inhibition of Plscr4 Induces a Hypertrophic Response in CMs

(A) The successful inhibition of Plscr4 was verified. The CMs were transfected with sh-Scramble or sh-Plscr4 for 48 hr (n = 6; *p < 0.05 versus sh-Scr). (B) Microscopic images of the CMs. Cells seeded in 24-well plates were cultured for 48 hr and then transfected with sh-Scramble or sh-Plscr4 for 48 hr. Both cell groups were then stained with antibodies directed against α-actinin and with DAPI for nuclear staining. Cell size was measured in 10 fields/well in both groups (n = 4 independent experiments; blue, nuclear; red, α-actinin; scale bars, 100 μm; n > 50 cells per experimental group; *p < 0.05 versus sh-Scr). (C) Protein/DNA ratio of the CMs. Cells were transfected with sh-Scramble or sh-Plscr4 for 48 hr. Then cells were lysed with standard sample buffer, protein concentration was determined by the BCA method with BSA as a standard, and DNA concentration was detected by fluorescence assay. The ratio of protein to DNA was then calculated to estimate potential protein synthesis (n = 6; *p < 0.05 versus sh-Scr). (D) The relative mRNA levels of the hypertrophic markers ANP, BNP, and β-MHC in the CMs treated as in (A) (n = 6; *p < 0.05 versus sh-Scr). (E) Representative western blot bands of β-MHC in the CMs that were transfected with sh-Scramble or sh-Plscr4 for 48 hr (n = 6; *p < 0.05 versus sh-Scr). All of the data are presented as the mean ± SEM.