Skip to main content
. Author manuscript; available in PMC: 2021 Jul 6.
Published in final edited form as: Curr Biol. 2020 Jun 4;30(13):2446–2458.e6. doi: 10.1016/j.cub.2020.04.057

Figure 3. Replacement of endogenous RLC with phosphomimetic/non-phosphorylatable mutants reveals a key role for Y155 in NMII assembly and adhesion maturation.

Figure 3.

(A) CHO.K1 cells were transfected with pSUPER-RLC or a control sequence (pSUPER-C), cultured for 96h and lysed. Lysates were separated by PAGE/SDS and blotted against RLC, MHCII-A and MHCII-B. Vinculin is shown as a loading control. Experiment is representative of eight performed.

(B) CHO.K1 cells were co-transfected with pSUPER-RLC together with GFP or RLC-GFP, wild type, Y155F or Y155E (first column). After 96h, cells were allowed to spread on fibronectin for 2h, fixed, permeabilized and stained as indicated. Bars=10 μm. Fourth column, magnification of the boxed regions in the vinculin column. Arrows point to adhesions. Arrowheads indicate the lack of adhesions in comparable regions.

(C-D) Quantification of the number of adhesions per μm2 (C) and adhesive area (D) in the conditions represented in (B). n ≥ 20 cells from three independent experiments (>2000 adhesions per condition). Significances are shown in the figures.

See also Figures S1 and S2.