VPS11 interacts with CORVET specific subunits.
A, MelJuSo cells co-expressing mRFP-RILP (red) and GFP-TGFBRAP1 or HA-VPS8 (green) were fixed, stained with antibodies against EEA1 (and anti-HA for VPS8), and imaged by CLSM. Scale bars: 10 μm. Graphs show average correlation coefficient (CC) ± S.E n > 25, for RILP or EEA1 and GFP, TGFBRAP1, or VPS8. Asterisks indicate significance to GFP control (****, p ≤ 0.0001; * p ≤ 0.05; ns = not significant). B, MelJuSo cells co-expressing GFP-TGFBRAP1 or HA-VPS8 were fixed, stained with antibodies against EEA1 (and anti-HA for VPS8), and imaged by CLSM. Scale bars: 10 μm. C, lysates of MelJuSo cells expressing GFP or GFP-VPS11 were immunoprecipitated (IP) with anti-GFP and analyzed by WB using anti-GFP and anti-TGFBRAP1 antibodies. Experimental conditions were run on the same blot with the same exposures for each detection antibody, and cutouts were taken and grouped for presentation purposes. TL, total lysate. D, MelJuSo cells expressing mRFP-VPS11 (blue) or co-expressing mRFP-VPS11 (blue) and GFP-TGFBRAP1 (green) were fixed, stained with antibodies against EEA1 (red), and imaged by CLSM. Scale bars: 10 μm. Graphs show average correlation coefficient (CC) ± S.E n > 25, between VPS11 and EEA1 ± ectopically expressed TGFBRAP1 (****, p ≤ 0.0001).