Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Enhanced large conductance K + channel activity contributes to the impaired myogenic response in the cerebral vasculature of Fawn Hooded Hypertensive rats

doi: 10.1152/ajpheart.00636.2013

Figure Lengend Snippet: Comparison of BK-α- and BK-β-subunit expression in cerebral vessel homogenates isolated from FHH and FHH.1BN rats. BK-α- and BK-β-subunits were probed on different blots. The same blots were then reprobed for β-actin. A: representative bands corresponding to the molecular weight of the BK-α-subunit (∼100 kDa), BK-β-subunit (∼25 kDa), and β-actin (∼42 kDa) in cerebral vessels obtained from FHH and FHH.1BN rats (n = 8 animals each; 3 sets of experiments). B: a comparison of the expression of α- and β-subunits. Numbers in parentheses indicate the number of samples studied.

Article Snippet: After transfer, the membrane was blocked with TBS-T buffer containing 20 mM Tris pH 7.5, 150 mM NaCl, 0.05% Tween, and a 5% blocking powder (Bio-Rad) at 4°C for 1 h. The blot was probed with primary antibody against BK α- and β-subunit [1:500 and 1:200, respectively; polyclonal rabbit Anti-K Ca 1.1, to amino acids 1184–1200 and Anti-sloβ1 (KCNMB1); Alomone Labs, Jerusalem, Israel] overnight at 4°C.

Techniques: Expressing, Isolation, Molecular Weight

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Magnesium isoglycyrrhizinate ameliorates radiation-induced pulmonary fibrosis by inhibiting fibroblast differentiation via the p38MAPK/Akt/Nox4 pathway.

doi: 10.1016/j.biopha.2019.108955

Figure Lengend Snippet: Fig. 4. MgIG regulated the expression of TGF-β1 and Nox4, and the phosphorylation of p38MAPK and Akt in vivo. The protein level of TGF-β1 in lung tissues at 12 weeks post-irradiation was determined using (A) im- munohistochemical staining (×200, n = 6) and (B) western blotting (n = 3), and quanti- tative analyses were performed. (C) TGF-β1 content in serum at 12 weeks post-irradiation was measured using an ELISA kit (n = 6). The protein levels of Nox4, and the phosphoryla- tion of p38MAPK and Akt in lung tissues at 12 weeks post-irradiation were observed using (D) immunohistochemical staining (×200, n = 6) and (E) western blotting (n = 3), and quanti- tative analyses were performed. All data were expressed as the mean ± SEM. #P < 0.05 vs. the control group; *P < 0.05 vs. the IR group; &P < 0.05 vs. the IR + MgIG group; Bar =50 μm.

Article Snippet: The serum was used to measure the TGF-β1 concentration using an ELISA kit (EK0515, Boster Bioengineering Institute, Huhan, China), according to the manufacturer's instructions.

Techniques: Expressing, Phospho-proteomics, In Vivo, Irradiation, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: MicroRNA-150 relieves vascular remodeling and fibrosis in hypoxia-induced pulmonary hypertension.

doi: 10.1016/j.biopha.2018.11.058

Figure Lengend Snippet: Fig. 4. Effect of miR-150 on pulmonary fibrosis of pulmonary hypertension rats. (A) Pulmonary fibrosis was detected by Masson’s staining (200× magnification). Scale bars, 100 μm. (B) The area of pulmonary fibrosis was calculated and shown. The mRNA expressions of TGF-β1 (C) and collagen I (D) in lung tissues were evaluated by qPCR. (E) The protein levels of TGF-β1 and collagen I in lung tissues were measured by western blot assay. (F)&(G) Relative grey values of the protein bands were shown. (H) The expressions of TGF-β1 and collagen I in lung tissues were detected by immunohistochemical staining (400× magnification). Scale bars, 50 μm. Data were presented as mean ± SD. **P < 0.01, ***P < 0.001 versus the indicated group.

Article Snippet: The membranes were blocked in 5% nonfat milk for 1 h at room temperature, then incubated with primary antibodies against TGF-β1 (1:400, BOSTER, China), Collagen I (1:300, BOSTER, China), p-AKTser473 (1:500, KeyGen, China), AKT (1:500, KeyGen, China), p-mTORser2481 (1:500, Sangon Biotech, China), mTOR (1:1000, Cell signaling Technology, USA), and β-actin (1:500, Bioss, China) at 4°C overnight, followed by incubation with HRP-labeled Goat AntiRabbit or Goat Anti-Mouse IgG (1:5000, Beyotime, China) at 37°C for 45min.

Techniques: Staining, Western Blot, Immunohistochemical staining

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: MicroRNA-150 relieves vascular remodeling and fibrosis in hypoxia-induced pulmonary hypertension.

doi: 10.1016/j.biopha.2018.11.058

Figure Lengend Snippet: Fig. 6. Effect of miR-150 on the expressions of fibrosis-related molecules. The mRNA expressions of TGF-β1 (A) and collagen I (B) in PASMCs were measured by qPCR. (C) The protein levels of TGF-β1 and collagen I in PASMCs were detected by western blot assay. (D)&(E) Relative grey values of the protein bands were shown. Data were presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 versus the indicated group.

Article Snippet: The membranes were blocked in 5% nonfat milk for 1 h at room temperature, then incubated with primary antibodies against TGF-β1 (1:400, BOSTER, China), Collagen I (1:300, BOSTER, China), p-AKTser473 (1:500, KeyGen, China), AKT (1:500, KeyGen, China), p-mTORser2481 (1:500, Sangon Biotech, China), mTOR (1:1000, Cell signaling Technology, USA), and β-actin (1:500, Bioss, China) at 4°C overnight, followed by incubation with HRP-labeled Goat AntiRabbit or Goat Anti-Mouse IgG (1:5000, Beyotime, China) at 37°C for 45min.

Techniques: Western Blot

Journal: Chinese Medicine

Article Title: Effects of solamargine in hepatic metastasis of colorectal cancer: induction of ferroptosis and elimination of cancer stem cells

doi: 10.1186/s13020-025-01171-5

Figure Lengend Snippet: SM impedes traits of CSCs partially through suppressing β-catenin in CRC. A Western blotting, B ALDH + staining and C tumorsphere formation assay were performed after CRC cells transduced with lentiviral β-catenin (encoded by the CTNNB1 gene) cDNA-expressing construct in the presence or absence of SM. Scale bar: 100 μm. D Western blotting, E ALDH + staining and F tumorsphere formation assays were conducted after CRC cells stably transduced with lentiviral shRNA against β-catenin with or without SM treatment. Scale bar: 100 μm. The values are represented as mean ± SD. * p < 0.05, *** p < 0.001

Article Snippet: Subsequently, the sections were incubated overnight at 4 °C with the primary antibodies against Ki-67 (Proteintech, 27309-1-AP), GSS (Proteintech, 15712-1-AP), GPX4 (Proteintech, 67763-1-Ig), Nrf2 (Proteintech, 16396-1-AP), β-catenin (Boster, PA1212), c-Myc (Abmart, T55150 ) and Cyclin D1 (Abcam, ab16663), followed by incubation with the secondary antibodies anti-mouse (ZSGB-BIO, PV-9002) and anti-rabbit (ZSGB-BIO, PV-9001).

Techniques: Western Blot, Staining, Tube Formation Assay, Transduction, Expressing, Construct, Stable Transfection, shRNA

Journal: Chinese Medicine

Article Title: Effects of solamargine in hepatic metastasis of colorectal cancer: induction of ferroptosis and elimination of cancer stem cells

doi: 10.1186/s13020-025-01171-5

Figure Lengend Snippet: SM inhibits hepatic metastasis of CRC cells partially by attenuating β-catenin. A Transwell migration and B matrigel invasion assays were conducted followed by CRC cells transduced with lentiviral β-catenin-encoding construct with or without SM treatment. Scale bar: 100 μm. C Transwell migration and D matrigel invasion assays were conducted followed by CRC cells transduced with lentiviral shRNA against β-catenin in the presence or absence of SM. Scale bar: 100 μm. E The overexpression efficiency of β-catenin in MC38-Luc cells was examined by Western blotting analysis. F Representative images and G quantitative analysis of photon flux on day 21 of C57BL/6 mice followed by intrasplenic injection of MC38-Luc cells stably express β-catenin. n = 5 per group. H Representative images and quantitative analysis of liver metastasis in C57BL/6 mice (n = 5) measured by bioluminescence imaging. I Representative images of metastatic livers and ( J ) the number of surface foci in the livers in C57BL/6 mice (n = 5) are shown. Arrows indicate the presence of metastatic nodules. K Representative photographs of H&E staining of the liver section and L the number of liver metastatic foci in microscopic fields are presented (n = 3). Scale bar: 200 μm. The values are represented as mean ± SD. * p < 0.05, *** p < 0.001

Article Snippet: Subsequently, the sections were incubated overnight at 4 °C with the primary antibodies against Ki-67 (Proteintech, 27309-1-AP), GSS (Proteintech, 15712-1-AP), GPX4 (Proteintech, 67763-1-Ig), Nrf2 (Proteintech, 16396-1-AP), β-catenin (Boster, PA1212), c-Myc (Abmart, T55150 ) and Cyclin D1 (Abcam, ab16663), followed by incubation with the secondary antibodies anti-mouse (ZSGB-BIO, PV-9002) and anti-rabbit (ZSGB-BIO, PV-9001).

Techniques: Migration, Transduction, Construct, shRNA, Over Expression, Western Blot, Injection, Stable Transfection, Imaging, Staining

Journal: iScience

Article Title: Kidney fibrosis molecular mechanisms Spp1 influences fibroblast activity through transforming growth factor beta smad signaling

doi: 10.1016/j.isci.2024.109839

Figure Lengend Snippet:

Article Snippet: TGF-β1 , Cell Signaling, USA , 5231LF.

Techniques: Recombinant, Staining, CCK-8 Assay, Enzyme-linked Immunosorbent Assay