Improving Protein Folding and Trafficking

To assess the ability of sub-physiological temperature incubation to improve protein folding and trafficking, cells expressing Cx30.3 or EKVP-linked mutants were incubated at 26°C for 48 h, fixed and subsequently immunolabelled as described earlier. To assess the potential for rescue of EKVP-linked mutants with chemical chaperones, connexin or mutant expressing cells were treated with 5 mM 4-phenylbutyrate (4-PBA; MilliporeSigma, Cat# 1716-12-7), or 500 μM tauroursodeoxycholic acid (TUDCA; MilliporeSigma, Cat# 14605-22-2). Drug treated cells were fixed and immunolabelled as described earlier. Connexin or mutant expressing cells treated with 10% (v/v) glycerol Caledon Laboratories, Georgetown, ON, Canada Cat# 5350-1) for 24 and 48 h were grown in glass-bottom 35 mm dishes and live-imaged as described earlier.