Extended Data Fig. 5: TMEM175-mediated H+ flux regulates juxta-lysosomal pH.
From: DNA nanodevices detect an acidic nanolayer on the lysosomal surface
a, Representative confocal images of TMEM175 KO HeLa cells transfected with D41A (a selective H+-conductance-deficient mutant with normal K+ permeability) or S45A (normal H+ conductance but with reduced K+-permeability), using CLMLY-SDNFAM/Cy5 (scale bar, 10 μm). Green cycles indicate transfected cells. Right panel shows the summary of juxta-lysosomal pH (n = 9 and 10 images for TMEM175 KO cells transfected with S45A and D41A, respectively). The red dashed line represents the juxta-lysosomal pH of TMEM175 KO HeLa cells. b, Arachidonic acid (ArA) induced juxta-lysosomal pH reduction in WT, but not TMEM175 KO HeLa cells. Representative images of CLMLY-SDNFAM/Cy5-engineered WT and TMEM175 KO HeLa cells treated with or without ArA (100 μM, scale bar, 10 μm). Right panel shows the summary of juxta-lysosomal pH (n = 8, 8, 9, and 7 images for the left to right groups, respectively). For all panels, 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 from 3 biological repeats. Two-tailed Student’s t test was used to assess statistical significance.
