Fig 1: PMA reverses the effect of ROCK2 silencing on apoptosis related protein expression in LPS-induced HK-2 cells. (A) Protein expression of Bcl-2, Bax, cleaved caspase 3/caspase 3 and cleaved PARP/PARP was measured using western blotting. (B) Protein expression levels of KIM-1 and NGAL was measured using western blotting. ***P<0.001 vs. control; $$$P<0.001 vs. LPS; #P<0.05, ##P<0.01 and ###P<0.001 vs. LPS + shRNA-ROCK2; +P<0.05, ++P<0.01 and +++P<0.001 vs. DMSO + LPS + shRNA-ROCK2. ROCK2, Rho-associated protein kinase 2; LPS, lipopolysaccharide; sh, short hairpin; PARP, poly (ADP-ribose) polymerase; KIM-1, kidney injury molecule-1; NGAL, neutrophil gelatinase-associated lipocalin; PMA, phorbol 12-myristate 13-acetate.
Fig 2: Mel reduced expression of kidney injury markers. (a) Immunofluorescence staining of NGAL (red). Magnification: 200x. (b) Immunofluorescence staining of KIM-1 (green). Magnification: 200x. (c) The expression of NGAL and KIM-1 was detected by western blot analysis. ****p < 0.0001 vs. control and ####p < 0.0001 vs. LPS, n = 3.
Fig 3: Patients with NPHPL and compound heterozygous mutations of XPNPEP3(A) Pedigrees of Family A and Family B. Family A harbors variants c.634G>C and c.761G>T; Family B harbors variants c.-87C>T and c.1261C>G. Arrows indicate nucleotide changes. Square for males; circles for females; solid for patients with NPHPL1; slash for deceased individuals.(B) Ultrasound image of patient A-II-1. Asterisks for medulla.(C) Abdominal CT image of patient B-II-1, showing the left kidney cyst (left panel) and left ilium cystic foci (right panel). Arrowheads indicate the foci.(D) Immunofluorescence analysis of KIM1 and NGAL in tubular epithelial cells. AQP1, aquaporin 1, a water channel protein located in the kidney proximal tubule. Bar: 5µm.(E) Schematic of XPNPEP3 gene variants. The XPNPEP3 gene is located on chromosome 22q. Both the c.634G>C and c.761G>T variants in Family A are located in exon 4, while the c.-87C>T and c.1261C>G variants in Family B are distributed in the noncoding region and exon 9, respectively.(F) Conservation analysis of XPNPEP3 proteins. Linear domain organization of the XPNPEP3 protein showing the p.A212 residue located in the AMP_N (aminopeptidase) domain and the p.R254 residue and p.H421 residue in the prolidase domain. MTS, mitochondrial target signal.
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