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. 2011 Jan 28;286(12):10834–10846. doi: 10.1074/jbc.M110.186668

FIGURE 5.

FIGURE 5.

Reduced munc18c expression does not contribute to syntaxin 4 knockdown phenotype. Syntaxin 4 was depleted in cells that inducibly express mouse FLAG-munc18c under control of the Tet-off system (FLAG-munc18c-syn4KD3 cells). A, expression levels of endogenous munc18c, FLAG-munc18c, and syntaxin 4 were determined by Western blotting. GAPDH was used as a loading control. Two different dox concentrations were used to regulate FLAG-munc18c expression. Please note the reduction of endogenous munc18c in the parental FLAG-munc18c cells when FLAG-munc18c is expressed. Also note that syntaxin 4 expression stays repressed in the presence of FLAG-munc18c. B, FLAG-munc18c and FLAG-munc18c-syn4KD3 cells were grown for 5 days on filter in the presence or absence of dox, fixed, and stained for claudin 2, Na,K-ATPase β1, and FLAG. This image shows a projection of x-y images. C, FLAG-munc18c and FLAG-munc18c-syn4KD3 cells were grown for 5 days on filter, fixed, and stained for Na,K-ATPase β1 and FLAG. This image shows a x-z scan. Scale bars, 20 μm.