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Fig. 4. PAR-3 knockdown decreases peripheral apical proteins in depolarized MDCK cells. (A) Western blot analysis of control cells (11-10 and 25-2) and PAR-3 kd cells (13-32 and 13-33) with antibodies of indicated proteins. Notice that the PAR-3 expression level was specifically reduced in clones 13-32 and 13-33. (B) A confluent monolayer of each cell on a permeable filter was incubated with LC medium for 20 hours, after which the cells were fixed and co-stained with anti-gp135 antibody and Rhodamine-phalloidin. (C) Cells treated as in B, stained with antibodies against ezrin and ZO-1. (D) Quantification of cells with cell peripheral gp135 staining. The percentage of total cells is shown. Values are mean (±s.d.) of three independent experiments (11-10 and 13-32) or mean of two independent experiments (13-33). Each photograph represents the projected views of optical sections from the apical to the basal membranes of cells. Bars, 20 µm.
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