Fig. 4: CALM depletion perturbs the intracellular localization of MT-RTKs.

a Parental, CALM iKD, and 18 h CPZ-treated MV4-11 cells were subjected to immunofluorescence microscopy. The cells were co-stained with the anti-FLT3 Ab (red), anti-CALM Ab (green), and 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bars, 20 µm. The right lower panels show regions with higher magnification. Images are representative of three independent experiments. b Parental, CALM iKD, and 18 h CPZ-treated MV4-11 cells were co-stained with the anti-FLT3 Ab (red), anti-protein disulfide isomerase (PDI) Ab (green), and DAPI (blue). Scale bars, 20 µm. The right lower panels show regions with higher magnification. Images are representative of three independent experiments. c The graph bars show the percentages of co-localization of FLT3 with PDI, Golgi matrix protein 130 (GM130), early endosome antigen-1 (EEA1), lysosome-associated membrane protein-1 (LAMP1), Rab11 and CALM. Results (%) represent the means ± SEM from ten cells. One-way ANOVA multiple comparisons with Bonferroni correction, *p < 0.0001. d Parental, CALM iKD, and 24 h CPZ-treated HMC-1 cells were subjected to immunofluorescence microscopy. The cells were co-stained with the anti-KIT Ab (red), anti-CALM (green) Ab, and DAPI (blue). Scale bars, 20 µm. Regions with higher magnification are shown in left lower panels. Images are representative of three independent experiments. e Parental, CALM iKD, and 24 h CPZ-treated HMC-1 cells were stained with the anti-KIT Ab (red), anti- LAMP1 Ab (green), and DAPI (blue). Scale bars, 20 µm. Regions with higher magnification are shown in left lower panels. Images are representative of three independent experiments. f The graph bars show the percentages of co-localization of KIT with PDI, GM130, EEA1, LAMP1, Rab11 and CALM with the means ± SEM from ten cells. One-way ANOVA multiple comparisons with Bonferroni correction, *p < 0.0001.