Fig. 4
Detection of LIMP-2 dimer in live cells using bimolecular fluorescence complementation method. a Schematic illustration of constructs designed for bimolecular fluorescence complementation in examining dimerization of LIMP-2. b High-resolution confocal images of Hela cells expressing LIMP-2 (WT, top, or H150A/F151D, bottom) fused to the N-terminal and C-terminal fragments of Venus fluorescent protein along with Lamp1-RFP co-transfected as a lysosome colocalization marker. c Western blot showing expression levels of the LIMP-2 WT and H150A/F151D variants using anti-myc and anti-HA antibodies. d Quantification of the ratio of mean Venus fluorescence intensity relative to ER-localized mCherry for cells transfected with LIMP-2 (WT and H150A/F151D) fused to the N-terminal and C-terminal fragments of Venus fluorescent protein fluorescence. Data is expressed as mean ± SEM, ~60 cells each experiment, n = 3, ****P < 0.0001, t test. Scale bar 20 µm
