BiFC analysis of the interactions of LMAN1 mutants with MCFD2 in living cells. (A) Fusion protein levels in cotransfection experiments were detected by immunoblotting (IB) using the indicated antibodies. Cells were lysed immediately after microscopic observations. (B) BiFC signals relative to the wild-type LMAN1 and MCFD2 fusion proteins. Fifty cotransfected cells from each set of transfection were counted, with the background subtracted for each image. Bars represent means ± SD from 3 replicates. Statistical analysis was performed using the Student t test, and asterisks indicate significant differences (P < .05).