Figure 4.
Figure 4. Septins regulate the lysosomal delivery of fluid-phase cargo. (A) MDCKs were transfected for 48 h with control or SEPT2 shRNAs and GFP or shRNA-resistant SEPT2-YFP (rescue). After a 5-min pulse/15-min chase with TR-dextran, cells were stained for LAMP1. Images show maximum-intensity projections of confocal image stacks. (B) Bar graph shows percentage (mean ± SEM) of TR-dextran particles that colocalize with LAMP1 (n = 20 cells). (C) Plot shows percentage (mean ± SEM) of TR-dextran with LAMP1 in control and SEPT2-depleted MDCKs (n = 15), which were pulsed with TR-dextran for 5 min and chased for the indicated times. (D) Lysates from MDCKs that stably express SEPT2-YFP at low and high levels were analyzed by SDS-PAGE and blotted for SEPT2. (E) Plot shows percentage (mean ± SEM) of TR-dextran with LAMP1 in MDCK cells (n = 14) that express SEPT2-YFP at low and high levels. Cells were pulsed with TR-dextran for 5 min and chased for the indicated times.

Septins regulate the lysosomal delivery of fluid-phase cargo. (A) MDCKs were transfected for 48 h with control or SEPT2 shRNAs and GFP or shRNA-resistant SEPT2-YFP (rescue). After a 5-min pulse/15-min chase with TR-dextran, cells were stained for LAMP1. Images show maximum-intensity projections of confocal image stacks. (B) Bar graph shows percentage (mean ± SEM) of TR-dextran particles that colocalize with LAMP1 (n = 20 cells). (C) Plot shows percentage (mean ± SEM) of TR-dextran with LAMP1 in control and SEPT2-depleted MDCKs (n = 15), which were pulsed with TR-dextran for 5 min and chased for the indicated times. (D) Lysates from MDCKs that stably express SEPT2-YFP at low and high levels were analyzed by SDS-PAGE and blotted for SEPT2. (E) Plot shows percentage (mean ± SEM) of TR-dextran with LAMP1 in MDCK cells (n = 14) that express SEPT2-YFP at low and high levels. Cells were pulsed with TR-dextran for 5 min and chased for the indicated times.

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