Figure 6.

NHE8 depletion causes an enlargement of acidic compartments. HeLa M-cells were treated with NHE8 siRNA using a double-knockdown protocol. Cells were treated with 30 μM DAMP for 20 min and then treated with or without 25 μM monensin (in the presence of DAMP) for a further 10 min before fixation. Cells treated with monensin are labeled “+ Mon” in the figure. Cells were immunolabeled with anti-LAMP1 antibodies (green) to label late endosomes/lysosomes and anti-DNP antibodies (magenta) to label the compartments containing DAMP. A series of confocal Z-stacks were taken with identical image settings and are displayed as an extended focus view. Enlarged areas are presented as inset boxes, and the arrows represent areas where the LAMP1 staining encircles the DNP staining. Scale bars, 17 μm.