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. 2017 Jan 15;350(2):336–348. doi: 10.1016/j.yexcr.2016.12.010

Fig. 3.

Fig. 3.

Knockdown of NMHC-IIA inhibits the interaction between VAMP2 and SNAP23. (A) Immunoblots showing VAMP2 in immunoprecipitates obtained with SNAP23 IgG but not with rabbit IgG from control or NMHC-IIA siRNA transfected mouse podocytes under starved (-insulin) or insulin-stimulated (+insulin) conditions. Control and NMHC-IIA siRNA-transfected podocyte lysates (30 µg) are included as controls. (B) Quantification of protein levels of three replicate blots as in (A) indicates that in insulin-stimulated mouse podocytes NMHC-IIA knockdown decreases complex formation between VAMP2 and SNAP23. (C–F) Duolink proximity ligation assay showing interaction between VAMP2 and SNAP23 in control siRNA (C, E) and NMHC-IIA siRNA (D, F) transfected cells. Each red spot represents an interaction detected by the kit. (G) Quantification of the spots in three replicate experiments as in (C–F) confirms the immunoprecipitation result showing that knockdown of NMHC-IIA fails to induce complex formation between VAMP2 and SNAP23. Scale bar, 30 µm. Bars show the mean and error bars STDEV of three independent experiments, Student's t-test. * p<0.05.