Fig. 7: Confirmation of possible FLOT1-/FLOT2-positive early endosome cargo proteins.
From: Cross-linking of the endolysosomal system reveals potential flotillin structures and cargo
a Co-IP of FLOT1 with possible cargo proteins detected by DIA-LC-MS/MS analysis of FLOT1-/FLOT2-positive early endosomes (n = 1). b Average DIA protein abundance for LPHN1, LPHN2, LPHN3, CLTA, EEA1, and RAB5C. Data are presented as mean values + SD (n = 3, biologically independent samples over three independent experiments). Significance based on Student’s unpaired two-sided t test; range: ∗P < 0.05, ∗∗P < 0.01; exact P values: LPHN1 (0.022), LPHN2 (0.004), LPHN3 (0.044), CLTA (0.047), EEA1 (0.035), RAB5C (0.008). c Co-IP of FLOT1 and FLOT2 with LPHN1, LPHN2, and LPHN3. LDLR serves as marker for plasma membrane and clathrin-mediated endocytosis. d Immunostaining of HeLa cells for FLOT1 and FLOT2 in combination with LPHN1, LPHN2, and LPHN3 (n = 3). Lower panels show a zoom-in for regions indicated in full-size images. Mander’s coefficients were determined to assess signal overlap between individual populations. Data are presented as mean values + SD (n = 3, cells examined over one experiment). Profile plots indicate the degree of colocalization for individual vesicles. Scale bars = 5 µM; Cntrl. control, IN input, WA wash, EL eluate. Source data are provided as a Source Data file.
