Fig 2: Delayed receptor-mediated endocytosis in ATP6ap2 deleted mice. WT/Pax8 + and Flox/Pax8 + mice were coinjected with dextran-FITC (10 kDa) and human recombinant transferrin. Kidneys were collected 10 and 40 min after injection. A Immunohistochemistry for dextran-FITC,10 kDa (green), DAPI (blue), and actin/phalloidin (red) in kidney slices from Wt and Flox/Pax8 + mice 10 and 40 min after injection. B Immunohistochemistry for human transferrin (red), DAPI (blue), and megalin (green) in kidneys from Wt and Flox/Pax8 + mice 10 and 40 min after injection. C Immunohistochemistry for Lamp-1 (green) and human transferrin (red) 40 min after injection. Scale bar size 50 µm for all pictures

Fig 3: Accumulation of autophagosomes in ATP6ap2 deficient mice. A, B Western blotting for total and phosphorylated mammalian target of rapamycin (mTOR) and the LC3B subunit of the autophagosome and densitometry summarizing results. Student’s t-test (n = 4 per group), **p = 0.01. C Immunohistochemistry for microtubule-associated proteins 1A/1B light chain 3B (LC3-B) (red) and DAPI (blue) in kidneys from Wt and Flox/Pax8 + mice insert shows higher magnification. Scale size bar 100 µm

Fig 4: Generation of kidney-epithelial cell-specific ATP6ap2 ablation in mouse. A RT-qPCR analysis of total mRNA of kidneys, lungs, and hearts from Wt and Flox/Pax8 + mice treated with 1 mg/mL doxycycline (low dose). Atp6ap2 mRNA abundance was normalized to HPRT. Statistical analysis was performed using Student’s t-test (Wt/Pax8 + : n = 4 and Flox/Pax8 + : n = 5) ***p < 0.001. B Western blotting for ATP6ap2 in total membrane preparations from the kidney of Wt and Flox/Pax8 + mice and summary of data as bar graph. Statistical analysis was performed using Student’s t-test (Wt/Pax8 + : n = 4 and Flox/Pax8 + : n = 5) *p < 0.05. C Immunohistochemistry for ATP6ap2 (red), AQP2 (green), and DAPI (blue) in kidney sections from Wt/Pax8 and Flox/Pax8 + mice with proximal tubules (upper panels) and medullary collecting ducts (lower panels). Scale bar size 100 µm

Fig 5: siPRR suppresses the pro-apoptotic ability of Ang II and miR-133a inhibitor. (A and B) The effects of siPRR on the levels of PRR expression in HUVECs treated with Ang II and miR133a inhibitor as measured by reverse transcription-quantitative polymerase chain reaction and western blotting. (C) Apoptosis of all HUVEC groups as determined by flow cytometry. Data are presented as the mean ± standard error of the mean (n=3). GAPDH was detected as an internal control. †P<0.05, ††P<0.01 vs. control group; §P<0.05, §§P<0.01 vs. NC group; #P<0.05, ##P<0.01 vs. NC + Ang II group; &P<0.05 vs. NC + Ang II + siPRR. Ang II, angiotensin II; miR, microRNA; NC, negative control; PRR, prorenin receptor.