FIGURE 4.

siRNA against hsc70 inhibits CaM-dependent nuclear import of SRY. A, immunoblot analysis (left panel) for the protein levels of hsc70 of HeLa cells treated with hsc70-specific small interfering RNA duplexes (hsc70 siRNA) 72 h post-treatment and probed with anti-hsc70 and anti-α/β tubulin antibody. Densitometric analysis (right panel) of images such as those in the left panel was performed to quantify the relative amount of hsc70 protein in the absence and presence of siRNA to hsc70 standardized relative to α/β-tubulin in whole cell extracts. Results are for a single typical experiment from a series of three similar experiments. B, HeLa cells were transiently transfected to express the indicated GFP-fusion proteins, 48 h post-treatment with siRNA to hsc70, and imaged live 20 h post-transfection by CLSM. C, results for the quantitative analysis, whereby CLSM images such as those in A were analyzed using ImageJ software for the nuclear to cytoplasmic ratio (Fn/c; fold extent of nuclear accumulation). Results are for the mean ± S.E. (n > 44) of a single typical experiment from a series of three similar experiments; p values are indicated where the Fn/c values are significantly different in the presence of hsc70 siRNA. WB, Western blot.