Fig. 5: CLIC5-F34D exhibits impaired membrane fusion capability.
From: Chloride intracellular channel (CLIC) proteins function as fusogens
a CLIC5-F34D shows reduced interaction with the membrane, as assessed by the co-floatation assay. Representative SDS-PAGE analysis and R18 fluorescence measurement are provided. Molecular weight markers in kDa are indicated. b Averaged fluorescence emission spectra following excitation at F280 for CLIC5-F34D (red) and CLIC5-WT (black; as in Fig. 1d) after incubation of 75 min. The peak at F344 corresponds to tryptophan emission, while the peak at F505 represents dansyl-PE emission due to the occurrence of FRET. c FRET ratio (F505/F344) comparison between CLIC5-WT and CLIC5-F34D, reflecting reduced membrane interaction of CLIC5-F34D. d–f CLIC5-F34D exhibits a markedly reduced effect on liposomal diameter as assessed by DLS (d), membranal lipid mixing monitored by R18 fluorescence unquenching (e), and carboxyfluorescein-mediated liposomal content mixing (f). For all experiments, data are presented as mean ± SEM, n = 3–4 independent experiments. Two-sided student’s t-test was performed for data analysis, ***P = 0.001.
