Related to Fig. 2 . SEC24C is needed for the association of RTN3L puncta with misfolded Akita. (A) Cells were transfected with LNPK-mCherry and Proinsulin-sfGFP and the percent proinsulin puncta that colocalize with LNPK was quantitated, bottom. Top, representative images are shown. Error bars represent SEM, n = 3 independent experiments. (B) siCtrl and siSEC24C-treated cells were transfected with Akita-sfGFP and mCherry-RTN3L and imaged. Arrowheads point to the Akita puncta that colocalize with RTN3L puncta. Right, quantification of the % Akita puncta positive for RTN3L puncta. (C) Percent cells with large Akita puncta (≥0.5 µm2) for the experiment shown in B. The large Akita puncta are marked with arrows in the images shown. Error bars represent SEM, n = 3 independent experiments. The results were quantified from 48 to 53 cells in A, 49–56 cells in B, and 113–125 cells in C. NS: not significant (P ≥ 0.05), * (P < 0.05), **(P < 0.01), Student’s unpaired t test.
