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. 2021 Apr 12;9:665919. doi: 10.3389/fcell.2021.665919

FIGURE 1.

FIGURE 1

The association of actin filaments with cytoplasmic CAV-1 in human osteosarcoma cells. (A) Top protein hits from the BioID screen for identification of Tpm3.1-associated proteins in U2OS cells. (B) Localization of endogenous CAV-1 and actin filaments in U2OS cells detected by CAV-1 antibody and fiuorescent phalloidin, respectively. Magnified regions from the area indicated by white boxes demonstrate that cytoplasmic CAV-1 aligns with actin filaments (white arrowheads). The bottom panels show the orthographic view of the enlarged region, where “XY” and “XZ” indicated different cross-sections. Bars, 10 μm (in cell images), 2 μm (in magnified images and orthographic views). (C) Time-lapse imaging of U2OS cells co-expressing CAV-1-mEGFP and mCherry-actin revealing that actin-associated cytoplasmic CAV-1 vesicles move along actin filaments (Box 1) and do retrograde flow with contractile arcs (Box 2). The magnified regions of Box 1 and 2 in different time point are shown on the right panels, respectively. White and yellow circles and dotted lines in the magnified regions indicate the starting and ending position of discrete CAV-1 tagged vesicles and contractile arcs, respectively. The recording was set as every 1 s for 200 s. Bars, 10 μm (in cell image) and 5 μm (in Box 1 and Box 2).