Fig. 10.

Effect of siRNA-mediated knock-down of Tmem147 expression in H508 cells on M3R-mediated increases in the expression of phosphorylated p90RSK (P-p90RSK). Cells were treated with negative control siRNA (Con) or Tmem147 siRNA (147) and grown in carbachol medium for 5 days, as described in detail under Materials and Methods. Cells were then acutely stimulated with 500 μM carbachol for 10 min. A, representative Western blot. B, summary of two or three separate immunoblotting experiments. P-p90RSK expression levels are expressed as means ± S.E.M. relative to the levels determined with cells that had been electroporated with negative control siRNA and had not been treated with 500 μM carbachol (100%; *, p < 0.05).