Fig. 7
TRAP depletion plus live-cell Ca2+ imaging reveals a TRAP function in Sec61-channel opening. HeLa cells were treated with the indicated siRNAs for 96 h, loaded with Fura 2, and subjected to live-cell imaging of cytosolic Ca2+ following our established procedure. Ca2+ release was unmasked by the addition of thapsigargin (TG) in the presence of external EGTA. a, c Average values are presented. Error bars represent standard error of the mean (s.e.m.). b, d Statistical analysis of the changes in cytosolic Ca2+ after TG addition in a, c. Error bars represent s.e.m. P-values of <0.001 by unpaired t-test were defined as significant, and are indicated by three asterisks (***). The numbers of analyzed cells are indicated. Data were collected in at least three independent experiments, with triplicate cultures for each condition. Knock-down efficiency was evaluated by western blots. e, f A representative western blot is shown, along with the silencing statistics (mean values with s.e.m.)
