Extended Data Fig. 4: Juxta-lysosomal pH of WT and TMEM175 KO cells determined using a lysosome-targeted, genetically-encoded pH indicator.
From: DNA nanodevices detect an acidic nanolayer on the lysosomal surface
a, Schematic illustration of the design of a genetically-encoded, lysosome-targeted, juxta-lysosomal pH probe, in which both pH-sensitive pHluorin and pH-insensitive mCherry are fused with lysosomal membrane protein TMEM192 at the cytosolic side (mCherry-TMEM192-pHluorin). b, Representative confocal images of WT and TMEM175 KO HeLa cells transfected with mCherry-TMEM192-pHluorin (scale bar, 10 μm). Right panel shows the summary of calibrated juxta-lysosomal pH values (n = 12 and 21 images for WT and TMEM175 KO cells, respectively). For panel b, statistical data are presented as box-and-whiskers graphs, where the box represents the range of 25%-75%, the solid line inside means the mean level, and the whiskers represent the minimum and maximum with n as randomly selected images/cells from 3 biological repeats. Two-tailed Student’ s t test was used to assess statistical significance.
