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. 2009 Dec 1;20(23):4962–4975. doi: 10.1091/mbc.E09-08-0712

Figure 9.

Figure 9.

Stable reexpression of mutants in the hydrophobic pocket of Munc18-1 largely restores syntaxin-1 expression and NE-secretion in the double knockdown clones (D7 and D16). (A) Munc18-1/-2 double knockdown clones (D7, D16) were infected with lentiviruses that express EmGFP, wild-type Munc18-1-EmGFP, or mutant (F115E, E132A. F115E/E132A) Munc18-1-EmGFPs and the infected cells were isolated with blasticidin. Thirty micrograms of total homogenates from heterologous pools of the surviving cells was analyzed by SDS-polyacrylamide gel electrophoresis and immunoblotting using anti-Munc18-1 and syntaxin-1 antibodies. Signals were detected with enhanced chemiluminescence detection system. A number on the left indicates the position of a molecular weight marker. (B and C) NE release was stimulated by 70 mM KCl for 15 min in the rescued cells (B for D7 clones; C for D16 clones). Error bars indicate SEM (n = 8 for D7 and n = 12 for D16).