Fig. 5: RAGA promotes CD47 lysosome localization.
From: RAGA prevents tumor immune evasion of LUAD by promoting CD47 lysosome degradation
a Immunofluorescence staining of CD47 and LAMP1 in control and RAGA knockdown A549 cells. The yellow arrow indicates cell surface CD47 staining. The white arrow indicates the colocalization of CD47 with LAMP1. b Statistical analysis of PM/intracellular CD47 expression ratio (n = 10 cells per group). c Statistical analysis of the percentage of CD47 colocalized with LAMP1 (n = 10 image areas per group). d Immunoblot analysis of indicated proteins in the isolated plasma membrane fraction of control and RAGA knockdown cells. e Immunoblot analysis of indicated proteins in the isolated lysosome fraction of control and RAGA knockdown cells. f Immunofluorescence staining of CD47 and RAB7 in control and RAGA knockdown A549 cells. The white arrow indicates the colocalization of CD47 with RAB7. g Statistical analysis of the percentage of CD47 colocalized with RAB7 (n = 10 image areas per group). h Construction of CD47 truncation mutants. i Immunoprecipitation analysis of the interaction between RAGA and CD47 truncation mutants. j Schematic model. RAGA interacts with CD47 TM and CT domains to promote the fusion of CD47-specific late endosomes with lysosomes, driving CD47 lysosome degradation. Loss of RAGA prevents the fusion of CD47 endosomes with lysosomes that causes the accumulation of late endosomes and increased CD47 levels. Data were from two independent experiments. Statistical data were presented as mean ± SD. P < 0.05 is considered statistical significance. ** indicates P < 0.01 and **** indicates P < 0.0001.
