Figure 5.

Dynasore moderately reduces CCh-induced M₂ endocytosis. (A) HEK 293T cells were transiently transfected with M₂-EGFP, starved for 18 h in serum-free medium and pretreated with 80 μM Dynasore or DMSO. Cells were left untreated, stimulated with 1 mM CCh for 15 min or incubated with Alexa Fluor 546-coupled transferrin (Tfn546, red); Cells were fixed, and the nuclei were stained with DAPI (shown in blue). Scale bars: 10 μm. (B) The number of cells displaying M₂ receptor internalization was quantified and is shown as the percentage of cells exhibiting intracellular M₂. At least 100 cells were counted per condition. Results are shown as the mean ± SD. Statistical analysis was performed with two-way ANOVA: *** p < 0.001; (C) Quantification of M₂-EGFP endocytosis in cytochalasin D treated cells.