Fig. 3. Endogenous Bpag1a/b2 stress fiber localization does not extend into focal contact sites, and most of the A' Ab staining occurs in and around the nucleus. (A-C) Observed by confocal microscopy, the A' antibody (A) stains in a pattern co-aligning perfectly with centrally-located stress fibers (stained with rhodamine-phalloidin in B; arrows in A and B). (D-F) In the same cells as in A-C, staining with the A' antibody (D) in an optical plane through the nuclei revealed strong nuclear (arrowheads) and perinuclear (arrow) staining (D). Rhodamine-phalloidin staining was absent from the nuclei (E). Hoechst staining was used to identify the nuclei (C,F). (G-I) When C2C12 cells were stained for both Bpag1a/b2 (G) and paxillin (H), and observed by confocal microscopy, the A' staining and the paxillin staining never overlapped (arrows in G and H). Shown is a region of a C2C12 cell with a large number of focal contact sites (paxillin staining) to illustrate the exclusion of Bpag1a/b2 from these sites (I is the merged image of G and H). Bars, 20 µm. Bar in A applies to A-F; bar in I applies to G-I.

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