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Fig. 7. Shrm alters the distribution of myosin II in MDCK cells. (A-C) Control (A), ShrmL-expressing (B) and Endo-ShrmC-expressing (C) cells were grown on filters and stained to detect non-muscle myosin II-B (green, A'-C', A'''-C''') and ZO-1 (red, A''-C''). Images are projections of 0.2 µm optical sections that span the AJC and apical plasma membrane. Areas boxed in A'-C' are shown enlarged in A'''-C'''. Lower panels represent X-Z projections of the entire apical-basal axis. Arrowheads denote where X-Z sections were generated. Arrows denote the regularly spaced distribution of non-muscle myosin II-B. (D,E) Mixed populations of wild-type T23 cells and ShrmL cells were grown on filters in the absence of doxycyclin and stained to detect myosin II-B (green, D' and E') and ZO-1 (red) or myosin IIB, ZO-1 and ShrmL (blue, E''). (F) Induced ShrmL cells were stained to detect myosin II-B (green) and F-actin (red). Arrowhead denotes position of X-Z projection shown beneath panel. F' and F'' show myosin llB and F-actin signals alone. (G) Induced ShrmL cells were stained to detect myosin IIB (green, G'), ShrmL (red, G'') and F-actin (blue, G'''). (H) Induced ShrmL cells grown on coverslips were stained to detect pp-RMLC (green, H'), ZO-1 (red, H'') and F-actin (blue, H'''). (I) Induced ShrmL cells were transiently transfected with a GFP--Actinin expressing vector and stained to detect myosin II-B (red, I') and GFP (green, I''). Scale bar equals 15 µm in A-H and 1.5 µm in I.

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