Figure 3: STK16 interacts with actin and regulates actin dynamics in cells in a kinase activity-dependent way.
From: STK16 regulates actin dynamics to control Golgi organization and cell cycle
(a) Immunoprecipitation shows that STK16 interacts with actin and both wild-type and the kinase-dead mutant STK16 bind to actin equally. HeLa cells that stably express STK16-GFP-FLAG-WT, STK16-GFP-FLAG-E202A or GFP-FLAG were subjected to immunoprecipitation with anti-Flag antibodies. Anti-Actin, anti-Tubulin and anti-FLAG antibodies were used in Western blots. The input/IP loading ratio was 1/10. (b) STK16 depletion causes actin depolymerization in vivo. HeLa, HeLa-STK16-GFP-FLAG-WT (WT) and HeLa-STK16-GFP-FLAG-MUT (RNAi resistant mutant, RNAi-X MUT) cells were transfected with control or STK16 siRNAs for 72 hours before they were fixed and stained for GFP (green) and fluorescence-labeled phalloidin (red). (c,d) STK16-IN-1 disrupts actin stress fiber earlier than Golgi complex. HeLa cells were treated with 10 μM STK16-IN-1 for 0, 15, 30, 60 minutes (not shown here), or 6 hours before they were harvested for immunofluorescence using fluorescence-labeled phalloidin (c) or anti-Giantin antibody (d). Experiments were repeated at least three times and representative results are shown. Scale bar: 10 μm.
