Fig. 2: TMEM9 and TMEM9B as obligatory binding partners of ClC-3, -4, and -5. | Nature Communications

Fig. 2: TMEM9 and TMEM9B as obligatory binding partners of ClC-3, -4, and -5.

From: Endosomal chloride/proton exchangers need inhibitory TMEM9 β-subunits for regulation and prevention of disease-causing overactivity

Fig. 2

a Distribution of T9A and T9B proteins in mouse tissues. Similar results obtained in two independent experiments. b T9 and CLC protein levels in brain and kidney of mice disrupted for Tmem9 (T9A) or Tmem9b (T9B). c T9, CLC, and marker protein levels in mouse embryonic fibroblasts from WT and T9a−/−/T9b−/− mice. d Clcn3 KO on ClC-4 and T9 protein levels. e Clcn4 KO on ClC-3 and T9 proteins. f Clcn5 KO on ClC-3 and T9 proteins. b–f, similar results observed in three independent experiments. g Clcn5 KO abolishes T9A and T9B expression, in green, in apical endosomes of renal proximal tubules (PT). Chimeric expression of X-chromosomal ClC−5 (left, red) in Clcn5+/− female mice reveals cell-intrinsic mechanism. h Specificity of T9 antibodies. Cortical sections of kidneys from T9a−/− and T9b−/− mice were stained with T9ACT and T9BCT antibodies against T9A and T9B. i T9B (green) does not colocalize with lysosomal ClC-7 (red) or lamp1 (red) in mouse renal PTs. Zoomed areas (white box) are displayed in lower left corner. j T9B (green) co-localizes with ClC-5 (red) on macropinosomes of BMDMs. Enlarged area is individually displayed for T9B and ClC-5. Similar results were obtained in two independent experiments. Scale bars, 10 µm throughout. Unprocessed blots are available in Source Data provided with this paper.

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