Skip to main content
. 2013 Mar 4;110(12):4610–4615. doi: 10.1073/pnas.1214887110

Fig. 3.

Fig. 3.

Stable reexpression of wild-type Munc18-1 or Munc18-2 restores the syntaxin-11 expressions and the degranulation defects of Munc18-1/2 DKD RBL-2H3 cells while functional specificities exist in PC12 cells. (A) Twenty micrograms of homogenates of Munc18-1/2 DKD PC12 cells rescued with EmGFP, wild-type Munc18-1, and Munc18-2 (without tag, with myc, and with EmGFP) were analyzed by SDS/PAGE and immunoblotting, using the antibodies indicated. (B) NA release was stimulated by 70 mM KCl for 15 min in the rescued cells. Error bars, SEM (n = 9). The statistical significance of the differences in NA release between wild-type Munc18-1 and Munc18-2 rescued PC12 cells in each tag configuration are indicated. *P < 0.05 (paired t test). (C) Munc18-1/2 DKD RBL-2H3 cells were infected with lentiviruses that express EmGFP, wild-type Munc18-1-EmGFP, and wild-type Munc18-2-EmGFP. Twenty micrograms of cell homogenates were analyzed by SDS/PAGE and immunoblotting, using the antibodies indicated. (D) β-Hexosaminidase release was stimulated by applying 0.01 μg/mL DNP-IgE and 50 ng/mL DNP-HSA, as well as 0.5 μM and 2.5 μM ionomycin for 1 h from wild-type RBL-2H3 cells and rescued cells. Error bars, SEM (n = 9). *Nonspecific band observed with rabbit polyclonal anti-Munc18-2 antibody that comigrated with Munc18-2-EmGFP.