Fig. 4.

Cisd2 interrupts Calnexin and SERCA complex formation. (A) HEK293T cells were transfected with V5-Calnexin (CNX), GFP-SERCA, GST, GST-Cisd2 constructs individually or in combinations. The expression level of GST-Cisd2 was varied between low (+) and high (++) levels. Two days post-transfection, whole cell lysates (WCL) were prepared and immunoprecipitated (IP) with anti-V5 antibodies. Co-immunoprecipitated proteins were detected by Western blotting by using anti-GFP, anti-GST, anti-V5, and anti-β-actin. (B) Representative Western blots of size-exclusion chromatography (SEC) fractions (13-25) from MEF cell lysates, prepared with or without Tg (0.3 μM for 6 h) treatment. Western blotting with anti-Calnexin (CNX), and anti-SERCA. (C) SEC fraction profiles represented as the percentage of Calnexin and SERCA in each fraction, indicating the amount per fraction relative to the total protein amount across all fractions. *P < 0.05, **P = 0.0094, ***P = 0.0009, by two-way ANOVA.