Extended Data Fig. 3: In vitro reconstitution of PDIA6 droplets.
From: A multichaperone condensate enhances protein folding in the endoplasmic reticulum
a, In vitro phase separation of PDIA6 (left) and Dylight 488–PDIA6 (right) with 10 mM Ca2+ in absence (top) and presence (bottom) of 10% PEG3350. Scale bar, 10 μm. b, In vitro time evolution of Dylight 488–PDIA6 droplets in presence of 10 mM Ca2+. Scale bar, 10 μm. c, FRAP experiments of Dylight 488–PDIA6 in presence of 10 mM Ca2+. Scale bar, 2 μm. d, Fluorescence recovery analysis of the Dylight 488–PDIA6 FRAP experiments presented in c. Experimental triplicate in shades of grey. e, Fusion event of Dylight 488–PDIA6 in presence of crowding agent and 10 mM Ca2+. Scale bar, 5 μm. f, In vitro phase separation of unlabelled PDIA6 in presence of crowding agent in absence (left) and presence of 10 mM Ca2+ (middle), and in presence of 10 mM Ca2+ and 20 mM EDTA (right). Scale bar, 10 μm. g, In vitro phase separation of Dylight 488–PDIA6 at different concentrations in presence of crowding agent and different Ca2+ concentrations after 30 min (left) and after 5 min (right), as indicated. Scale bar, 10 μm.
