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. Author manuscript; available in PMC: 2023 Jun 20.
Published in final edited form as: Curr Biol. 2022 May 19;32(12):2704–2718.e6. doi: 10.1016/j.cub.2022.04.088

Figure 4: MARK2 association with the plasma membrane is required for leading edge advance and FA localization.

Figure 4:

Imaging and analyses of U2OS cells (U2OS), or U2OS cells with MARK2 knocked out by CRSPR-cas9 (MARK2-KO) and MARK3 knocked down by siRNA (MARK-KO) or MARK2-KO cells expressing MARK2-GFP or MARK2-GFP lacking amino acids 1098-1192 (MARK2-ΔKAGFP). (A) Diagram of domains for MARK2 and MARK2-ΔKA lacking the KA1 membrane-targeting domain. (B) Time-lapse image series (time in min) and kymograph (B’) taken along the red line (right panel distance (d) and time (t)) of a cell expressing MARK2-GFP plated on a “crossbow”-shaped fibronectin island surrounded by non-adhesive substrate. Bar=10μm. (C-E) Localization of MARK2-GFP (C, D top, E) or MARK2-ΔKA-GFP (C, D, bottom, F) and staining of F-actin (C-F), paxillin (E, F) or tubulin (E, F) in fixed (C) or pre-extracted and fixed cells (D, E, F). White boxes: regions magnified at right (zoom), white lines: regions used for line scan analyses in (E’, F’). Circle and arrows in (C, D: MARK2 at the centrosome and a protrusion, respectively. Bars in (C, D, and E (left)= 5μm, (E, right)= 2μm. See also Figure S4, Video S5.