Figure 4
From: Mahogunin regulates fusion between amphisomes/MVBs and lysosomes via ubiquitination of TSG101
Functional aberration of MGRN1 blocks fusion of vesicles with lysosomes. (a) HeLa cells treated with mock or MGRN1 siRNAs and transiently transfected with RFP-LC3 construct were subjected to Alexa-Fluor 488 EGF uptake. Cells were washed, fixed at indicated time points and imaged. Insets reveal enlarged views of green (Alexa-Fluor 488-labeled EGF), red (RFP-LC3) and yellow (Alexa-Fluor 488 EGF and RFP-LC3 colocalized) vesicles as shown by respective colored arrow heads. Scale bar, 5 μm. (b) Graph represents average number of red, green and yellow fluorescent vesicles analyzed over five fields. Error bars, ±S.E.M. (c) SHSY5Y cells transiently transfected with MGRN1 or MGRNΔR and RFP-LC3 construct were subjected to Alexa-Fluor 488 EGF uptake in a similar experiment as (a). The time points at which fixed cells were imaged are indicated. Insets reveal enlarged views of green (Alexa-Fluor 488-labeled EGF), red (RFP-LC3) and yellow (Alexa-Fluor 488 EGF and RFP-LC3 colocalized) vesicles as shown by respective colored arrow heads. Scale bar, 5 μm
