Fig 17. Effect of knockdown and inhibition of ROCK1 and ZIPK on LC₂₀ phosphorylation under serum-free conditions assessed by Phos-tag SDS-PAGE.

(A) ROCK1 or ZIPK was down-regulated by siRNA treatment. Controls were treated identically, but with negative control siRNA. Cells were serum starved overnight prior to lysis in Laemmli sample buffer for Phos-tag SDS-PAGE and western blotting with anti-LC₂₀. A representative western blot is shown in the left-hand panel with cumulative quantitative data on LC₂₀ phosphorylation stoichiometry in the right-hand panel. Statistical analysis was carried out with Dunnett’s post hoc test. (B) The effect of inhibition of ROCK or ZIPK on LC₂₀ phosphorylation in UASMC assessed by Phos-tag SDS-PAGE and western blotting with anti-LC₂₀. A representative western blot is shown in the left-hand panel with cumulative quantitative data on LC₂₀ phosphorylation stoichiometry in the right-hand panel. Statistical analysis was carried out with Dunnett’s post hoc test.