FIG. 3.

Dyn-2 and Cav-1 expression in HepaRG cell lines is stable during differentiation. HepaRG^Dyn-2, HepaRG^Dyn-2K44A, HepaRG^Cav-1, and HepaRG^Cav-1Δ1-81 cells were differentiated in the presence of 1.8% DMSO or maintained undifferentiated by splitting them at 2-day intervals in normal growing medium as controls. (A) Undifferentiated (lanes 1 and 3) and differentiated (lanes 2 and 4) HepaRG^Dyn-2 and HepaRG^Dyn-2K44A cells were lysed, and equal amounts of protein were subjected to SDS-PAGE, followed by Western blotting analysis of endogenous and overexpressed Dyn-2 (marked as Dyn-2 and Dyn-2 EGFP, respectively) using anti-Dyn-2 Ab. The contaminating band, marked with an asterisk and usually present in retrovirus-infected HepaRG cells, was used as a loading control. (B) Undifferentiated (black bars) and differentiated (open bars) HepaRG^Cav-1 and HepaRG^Cav-1Δ1-81 cells were lysed, and total RNA was purified. Cav-1 expression was quantified by real-time RT-PCR using the Cav-1-FL primers. Amplification of β-actin in the same samples was used as a loading control. The results were expressed as Cav-1 fluorescence units (FU) divided by β-actin FU.