Fig. 2.

A: PSCs in the control group had minimal transient receptor potential vanilliod type 4 (TRPV4) immunoreactivity (green) (n = 145). 4,6-Diamidino-2-phenylindole (DAPI, blue) nuclear stain is shown. B: TRPV4 expression increased in the cytoplasm (n = 108) of PSCs after exposure to EtOH (50 mM) for 24 h. C: histogram shows the average fluorescent intensity of TRPV4 in alcohol-treated PSCs was significantly increased (***P < 0.001, Student's t-test). D: Western immunoblot analysis was used to compare TRPV4 protein in PSCs pooled from rats chronically fed an alcohol and high-fat diet (HFA, 6%, 5–6 wk; n = 4) with TRPV4 in PSC from control rats fed regular rodent chow (n = 4). TRPV4 expression was increased in the HFA-fed group. E: bar graph summarizes the relative density of the TRPV4 expression in immunoblot bands for both HFA-fed and control groups fed normal chow normalized with β-actin. There was approximately a 20% increase in expression for the PSCs isolated from HFA-fed rats over cells from control rats fed a normal diet. (*P < 0.05). F: RT-PCR was performed with primary cultured PSCs from control and HFA-fed rats. The predicted PCR amplicon sizes for TRPV4 is 339 bp. The PCR marker contains a 50–2000 bp ladder (Invitrogen, Grand Island, NY).