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. 2009 Nov 24;122(24):4465–4472. doi: 10.1242/jcs.055228

Fig. 3.

Fig. 3.

EDEM1 knockdown results in the accumulation of P23H rod opsin. P23H rod opsin-GFP (P23H-GFP) and empty vector (A) or P23H-GFP and shEDEM1 (B) were transfected in SK-N-SH cells. 24 hours after transfection, the cells were fixed and analysed by confocal microscopy. Scanning parameters during image acquisition were kept constant. Arrows indicate increased ER staining and inclusion formation. Scale bar: 10 μm. (C) The incidence of inclusion formation of P23H-GFP was assessed with increasing shEDEM1 plasmid (1:1, 1:2, 1:3, 1:4). Error bars represent ± 2 s.e.m. *P=0.0134. (D) Constant pMT3-P23H rod opsin and increasing (1:2, 1:3, 1:4) molar plasmid equivalents of shEDEM1 were transfected in SK-N-SH cells. DM-soluble fractions were obtained 24 hours after transfection and 10 μg protein resolved by SDS-PAGE. Asterisk indicates species showing greatest accumulation. Rod opsin was detected using 1D4 mAb. The position of molecular size markers in kDa are indicated on the left.