FHL2 slows down the turnover of Arkadia.
A and B, 293T cells were transfected with Flag-tagged Arkadia with or without FHL2 and treated with cycloheximide (CHX) as indicated. Immunoblotting analysis was performed with anti-Flag antibody (A). Signal intensities were plotted relative to 0 h values. Data are presented as the mean values ± S.D. from three independent experiments (B). C and D, 293T cells were transfected with increasing doses of FHL2 (+: 0.5 μg, ++: 1.0 μg, +++: 1.5 μg). Endogenous Arkadia was analyzed by immunoblotting (C). Signal intensities were plotted relative to the values without transfected FHL2. Data are presented as the mean values ± standard deviation from three independent experiments (D). E and F, knockdown of FHL2 in HaCaT cell. HaCaT cells were transduced with either control (Ctl) or two independent FHL2 shRNA (shFHL2) lentiviral particles (5774 and 5773). Expression of endogenous Arkadia was analyzed by immunoblotting (E). Signal intensities were measured relative to the values transduced with control particles (F).