Fig. 5.

ALIX and the ESCRT-I recruitment factor TSG101 are corecruited with ALG-2 to damaged lysosomes. (A) U2OS cells were treated with either DMSO (Upper row) or LLOME (Lower row) for 15 min and then immunostained for ALG-2 (green), ALIX (magenta), TSG101 (yellow), and Gal-3 (cyan). Relatively large colocalized puncta of ALG-2, ALIX, and TSG101 along with Gal-3 were observed after LLOME treatment. The regions encapsulated within the white boxes have been enlarged and the individual protein channels are shown on the Right. Images are representative of five independent replicates. (B) The area of overlapping compartments increased dramatically upon LLOME treatment compared to the DMSO control (n = 75 cells for DMSO, 64 cells for LLOME). The circles on the bar charts represent independent data points and the data are shown as mean ± SD (vertical line). P = 0.0002 (***). (Scale bar, 5 μm.)