Digital images and densitometry for Western blot analysis of TRPV4 from total lysate (A) and membrane fractions (B) of 4B cells. In A, cells were incubated with control media or 100 nM ANG II for 1 h, and densitometric analysis of TRPV4 immunoreactivity was performed using GAPDH for normalization. In B, membrane fractions were isolated from 4B cells incubated for 1 h in control media of 4B cells or with 100 nM ANG II alone and in combination with 1 μM losartan, 10 μM PP2, or 10 μM PP3. Densitometric analysis of TRPV4 immunoreactivity was normalized with Flotillin. Data are expressed as means ± SE. Numbers of independent experiments are provided in parenthesis. *Significantly different from Control, Los + ANG II, and PP2 + ANG II (P < 0.05, ANOVA followed by Student-Neuman Keuls post hoc analysis), **Significantly different from ANG II and PP3 + ANG II (P < 0.05, ANOVA followed by Student-Neuman Keuls post hoc analysis).