Fig. 4

Effects of single-point mutations in DxxxLI motif on FACI’s localization to CCPs. A Immunoprecipitation (IP). HepG2 cells were transfected with mEmerald-FACI or mEmerald-FACI-ΔDxxxLI plasmid. Cells were lysed and immune-precipitated with anti-GFP (i.e., anti-mEmerald). Immunoprecipitates were analyzed by SDS-PAGE and probed with anti-GFP and anti-AP2M1 antibodies. B DxxxLI motif of FACI proteins was mutated point by point. Amino acid sequences of WT and a series of “DxxxLI”-mutated FACI were shown. C TIRFM images. AML12 cells expressing mCherry-AP2M1 (left) or mCherry-CLC (right) were transfected with WT or “DxxxLI”-mutated mEmerald-FACI construct. Scale bar, 5 µm