Extended Data Fig. 2: Effect of ER stress on PDIA6 condensates.
From: A multichaperone condensate enhances protein folding in the endoplasmic reticulum
a, Fixed HeLa cells co-transfected with PDIA6–GFP (pink) and Akita proinsulin-myc (green). Akita proinsulin, a folding-defective mutant of proinsulin18, was used to mimic the accumulation unfolded/misfolded proteins in the ER lumen. Akita proinsulin forms puncta, as previously reported61. PDIA6 was visualized by imaging eGFP, Akita proinsulin using an anti-myc antibody. Scale bar, 20 μm, insert 5 μm. b, Western blot showing expression levels of PDIA6, BiP and α-tubulin and at indicated time points of Tm treatment in HeLa cells. Unprocessed blots are available. Representative blot of three independent experiments. c, Quantification of PDIA6 and BiP levels from blot shown in b. Data representative of an experimental triplicate (n = 3). Mean and standard deviation. d–f, Fixed HeLa cells expressing endogenous PDIA6 and live-cell imaging of HeLa cells transfected with PDIA6–GFP after 10 h of Tm treatment (d), 10 h of Tg treatment (e) or 16 h of CPA (reversible Tg analogue) treatment (f) (left) and 24 h after drug washout (right). Endogenous PDIA6 was stained with an anti-PDIA6 antibody, overexpressed PDIA6–GFP was visualized by imaging eGFP. g, Fixed HeLa cells expressing endogenous PDIA6 observed at different time points after Tm (top) and Tg treatment (bottom).
