Fig 1: BMPER inhibited Erk1/2 phosphorylation in NRK-49F cells. NRK-49F cells were incubated with 2 ng/ml TGF-β1 and increasing amounts of BMPER as indicated for 20 min, and Erk1/2, p- Erk1/2 were measured by Western blot (A). The relative protein expression for p-Erk1/2 / Erk1/2 in NRK-49F cells was displayed (B). NRK-49F cells were treated with TGF-β1 (2 ng/ml) or TGF-β1 (2 ng/ml) plus BMPER (80 nM) at indicated time, and Erk1/2, p- Erk1/2 were measured by Western blot (C). The relative protein expression for p-Erk1/2 / Erk1/2 in NRK-49F cells was displayed (D). Values are the means ± SEM (n = 3). *p < 0.05 vs. TGF-β1 group.
Fig 2: BMPER ameliorated tubule atrophy and interstitial fibrosis. H&E staining showed BMPER ameliorated tubular injury [upper panel of (A)]. Arrows indicated tubular atrophy. PSR staining showed BMPER decreased interstitial fibrosis [lower panel of (A)]. Semi-quantitative analysis for tubule atrophy and interstitial fibrosis were shown in (B,C), respectively. Scale bar = 100 μm. Values are the means ± SEM (n = 6). *p < 0.05 vs. sham; †p < 0.05 vs. UUO+pcDNA3.
Fig 3: The inhibitory effect of BMPER on Erk1/2 phosphorylation was dependent of LRP1 in NRK-49F cells. The effect of LRP1 siRNA transfection on LRP1 protein expression (A) Erk1/2, p- Erk1/2 were measured by Western blot 15 min after NRK-49F cells with different treatments (B). The relative protein expression for α-SMA and p-Erk1/2/Erk1/2 in NRK-49F cells was displayed (C,D). Values are the means ± SEM (n = 3). *p < 0.05 vs. cells treated with TGF-β1 plus BMPER.
Fig 4: Exogenous BMPER inhibited TGF-β1-induced dedifferentiation in HK-2 cells. HK-2 cells were incubated with increasing amounts of BMPER (5–80 nmol), and cell viability was detected by MTT (A). HK-2 cells were treated with 10 ng/ml TGF-β1 and increasing amounts of BMPER as indicated for 48 h, and the protein expression for Id1, E-cadherin and α-SMA was measured by Western blot (B). The relative protein expression for Id1, E-cadherin and α-SMA in HK-2 cells was displayed (C–E). Values are the means ± SEM (n = 3). Representative photographs for E-cadherin in HK-2 cells after various treatments were displayed by immunofluorescence staining (F). Graphic presentation of mean fluorescent intensity in various groups (G). Neither treatment modalities significantly affected HK-2 cell counts (H). Cell numbers were counted after various treatments for 48 h. Scale bar = 100 μm. *p < 0.05 vs. control; †p < 0.05 vs. TGF-β1 group.
Fig 5: BMPER expression decreased in kidneys after UUO and in HK-2 cells exposed to TGF-β1. Kidneys with indicated UUO time and sham controls were subjected to immunohistochemical staining for BMPER (A). Arrows indicated BMPER-positive tubular cells. Scale bar = 100 μm. BMPER expression in kidneys was measured by Western blot (B). The relative protein expression for BMPER in kidneys was displayed (C). The mRNA expression of BMPER in kidneys with sham operation and UUO was shown (D). Values are the means ± SEM (n = 6). HK-2 cells were exposed to TGF-β1 at indicated time and BMPER expression was measured by Western blot (E). The relative protein expression for BMPER in HK-2 cells was displayed (F). The mRNA expression of BMPER in HK-2 cells was shown (G). HK-2 cells were exposed to TGF-β1 at various concentration and BMPER expression was measured by Western blot (H). The relative protein expression for BMPER in HK-2 cells was displayed (I). The mRNA expression of BMPER in HK-2 cells was shown (J). Values are the means ± SEM (n = 3). †p < 0.05 vs. sham; *p < 0.05 vs. control.
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