tgf β1 receptor i  (Abcam)

Journal: Brazilian Journal of Pharmaceutical Sciences

Article Title: Blockage of TGF-β1-induced epithelial-to- mesenchymal transition by oxymatrine prevents renal interstitial fibrosis

doi: 10.1590/s2175-97902020000118738

Figure Lengend Snippet: FIGURE 4 - TGF-β1 protein levels were determined by ELISA in the kidneys at 3d or 7 d after UUO with or without OM treatment and sham group The protein levels of TGF-β1 type I receptor at 3 d or 7 d after UUO with or without OM. OM reduced the TGF-β1 and its type I receptor (TGF-βRI) levels of renal tissues in UUO mice. The values are represented as the density of TGF-βRI vs tubulin control. The graph summarizes densitometric analysis of 3 independent technical experiments. *: P<0.05 vs. sham group, **: P<0.05 vs. UUO-3d and UUO-7d group respectively.

Article Snippet: Monoclonal antibodies against FN (sc-71113), TGF-β1 type I receptor (TGFβRI) (sc-398) and α-tubulin (sc-58667) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Brazilian Journal of Pharmaceutical Sciences

Article Title: Blockage of TGF-β1-induced epithelial-to- mesenchymal transition by oxymatrine prevents renal interstitial fibrosis

doi: 10.1590/s2175-97902020000118738

Figure Lengend Snippet: FIGURE 5 - The dose response effect of OM on proliferation of HK-2 cells At the concentration of 60- 360 mg/L, OM increased cell viability in a dose-dependent manner, while above 360 mg/L this effect is not obvious although there was no statistically significant difference (A). OM blocks TGF-β1-mediated EMT in vitro. HK-2 cells were treated without (control) or with 5 ng/ml TGF-β1 in the presence or absence of various concentrations of OM as indicated for 48 H (B and C). Western blot showed that OM restored E-cadherin expression that was inhibited by TGF-β1 in a dose-dependent manner, whereas it suppressed TGF-β1-mediated induction of FN and α-SMA. Whole-cell lysates were immunoblotted with antibodies against FN, α-SMA, E-cadherin and tubulin, respectively. Densitometric analysis of three independent technical experiments (B and C). The data were normalized by the density of tubulin control. *: P<0.05 vs. control group, **: P<0.05 vs. TGF-β1 group, #: P<0.05 vs. TGF-β1 group, respectively.

Article Snippet: Monoclonal antibodies against FN (sc-71113), TGF-β1 type I receptor (TGFβRI) (sc-398) and α-tubulin (sc-58667) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Concentration Assay, In Vitro, Control, Western Blot, Expressing

Journal: Brazilian Journal of Pharmaceutical Sciences

Article Title: Blockage of TGF-β1-induced epithelial-to- mesenchymal transition by oxymatrine prevents renal interstitial fibrosis

doi: 10.1590/s2175-97902020000118738

Figure Lengend Snippet: FIGURE 6 - Immunofluorescence staining demonstrated that OM blocks TGF-β1-mediated EMT in HK-2 cells Immunofluorescent microscopy of FN (A through I), α-SMA (J through R) and E-cadherin (S through &) in HK-2 cells after various treatments. HK-2 cells were treated without (A, J, S) or with TGF-β1 at 5 ng/mL (B, K, T), or TGF-β1 plus 360 mg/L of OM (C, L, U) for 48 h. Basline FN (A, G), α-SMA (J, P) and E-cadherin (S, Y) in HK-2 cells. FN (B, H) and α-SMA (K, Q) upregulated and E-cadherin (T, Z) downergulated after TGF-β1 treatment. The increased immunolabeling intensity of FN, α-SMA and the decreased intensity of E-cadherin in response to TGF-β1 stimuli were reversed by OM treatment (C and I, L and R, U and &). Nuclear staining for DAPI is shown (D- F, M- O and V- X) and merged results are also demonstrated (G- I, P- R and Y- &). The graph summarizes densitometric analysis of 3 independent technical experiments. Magnification A- &: ×400. OM treatment attenuated the expression of TGF-βRI in HK-2 cells activated by TGF-β1

Article Snippet: Monoclonal antibodies against FN (sc-71113), TGF-β1 type I receptor (TGFβRI) (sc-398) and α-tubulin (sc-58667) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Immunofluorescence, Staining, Microscopy, Immunolabeling, Expressing

Journal: Brazilian Journal of Pharmaceutical Sciences

Article Title: Blockage of TGF-β1-induced epithelial-to- mesenchymal transition by oxymatrine prevents renal interstitial fibrosis

doi: 10.1590/s2175-97902020000118738

Figure Lengend Snippet: FIGURE 7 - OM treatment attenuated the expression of TGF-βRI in HK-2 cells activated by TGF-β1 HK-2 cells were incubated with TGF-β1 (5 ng/ml) or/and various concentrations of OM for 48 H. The protein expression of TGF-βRI in HK-2 cells was significantly increased after incubation with TGF-β1 (5 ng/ml), while it was inhibited by OM in a dose-dependent manner (A). Graphical presentation of the relative abundance of TGF-βRI after normalization with α-tubulin control in different groups. The graph summarizes densitometric analysis of 3 independent technical experiments. *: P<0.05 vs. control group, **: P<0.05 vs. TGF-β1 group, #: P<0.01 vs. TGF-β1 group.

Article Snippet: Monoclonal antibodies against FN (sc-71113), TGF-β1 type I receptor (TGFβRI) (sc-398) and α-tubulin (sc-58667) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Incubation, Control

Journal: International Journal of Biological Sciences

Article Title: Anp32e promotes renal interstitial fibrosis by upregulating the expression of fibrosis-related proteins

doi: 10.7150/ijbs.74431

Figure Lengend Snippet: Anp32e activated the TGF-β1/Smad3 pathway in BUMPT cells. (A) ELISA analysis of TGF-β1 levels in BUMPT cells transfected with empty vector or Exp-Anp32e plasmids (n=3). (B-D) Western blotting and densitometry analysis of TGF-β1, t-Smad3 and p-Smad3 proteins in BUMPT cells transfected with empty vector or Exp-Anp32e plasmids (n=3). (E-H) Western blotting and densitometry analysis of t-Smad3, p-Smad3, Fn, and Col-I proteins in BUMPT cells transfected with empty vector or Exp-Anp32e plasmids, treated with or without SB431542 (n=3). * P < 0.05, ** P <0.01 vs . empty vector group, # P < 0.05 vs . Exp-Anp32e group. Data represent means ± SD.

Article Snippet: Subsequently, to inhibit the TGF-β1/Smad3 signaling pathway, BUMPT cells were treated with 10μM type I TGF-β1 receptor inhibitor SB431542 for 24 hours after transfection with Anp32e overexpression plasmids for 12 hours. (APExBIO, Houston, TX, United States).

Techniques: Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Western Blot