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: Cell Communication and Signaling : CCS

Article Title: SENP1 regulates the transformation of lung resident mesenchymal stem cells and is associated with idiopathic pulmonary fibrosis progression

doi: 10.1186/s12964-022-00921-4

Figure Lengend Snippet: The expression of SENP1 increases in LR-MSCs from mice. A LR-MSC morphology was observed using light microscopy after mouse LR-MSCs were treated with TGFβ1 (10 ng/mL) for 7 and 14 d. Black arrows indicate LR-MSCs. B Representative images of dual staining for Collagen I (green) and α-SMA (red), SENP1 (green) and α-SMA (red), and dual staining for SENP1 (red) and Collagen I (green). C Western blotting was applied to detect the levels of α-SMA, SENP1, Collagen I in LR-MSCs after the TGFβ1 (10 ng/mL) treatment for 14d. Scale bar, 20 μm. * P < 0.05, control group vs. the TGFβ1 7d group; ** P < 0.01, control group vs. the TGFβ1 14d group. The results are shown as the means ± SD. Each experiment was performed in triplicate

Article Snippet: Recombinant TGFβ1 (Proteintech Group, Wuhan, China) was used at a 10 ng/mL concentration to induce LR-MSC myofibroblast transition.

Techniques: Expressing, Light Microscopy, Staining, Western Blot, Control

Journal: Cell Communication and Signaling : CCS

Article Title: SENP1 regulates the transformation of lung resident mesenchymal stem cells and is associated with idiopathic pulmonary fibrosis progression

doi: 10.1186/s12964-022-00921-4

Figure Lengend Snippet: The LR-MSCs-myofibroblast transition was inhibited upon SENP1 downregulation in vitro and in vivo. A Representative images of dual staining in vitro for Collagen I (green) and α-SMA (red), SENP1 (green) and α-SMA (red), and dual staining for SENP1 (red) and Collagen I (green). Scale bar, 15 μm. B Collagen I, α-SMA, and SENP1 levels were assessed using western blotting analyses. Quantification is shown in the lower panel. C Representative images of dual staining in vivo for Sca-1 (green) and α-SMA (red), Sca-1 (green) and SENP1 (red). Scale bar, 50 μm. * P < 0.05, ** P < 0.05, # P < 0.01, ## P < 0.01. *, control group vs . the TGFβ1 7d group. #, control group vs . the TGFβ1 14d group. **, the SENP1 knockdown group vs . the TGFβ1 7d group. ##, the SENP1 knockdown group vs . the TGFβ1 14d group. Each experiment was performed in triplicate

Article Snippet: Recombinant TGFβ1 (Proteintech Group, Wuhan, China) was used at a 10 ng/mL concentration to induce LR-MSC myofibroblast transition.

Techniques: In Vitro, In Vivo, Staining, Western Blot, Control, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: SENP1 regulates the transformation of lung resident mesenchymal stem cells and is associated with idiopathic pulmonary fibrosis progression

doi: 10.1186/s12964-022-00921-4

Figure Lengend Snippet: SENP1 regulates LR-MSC myofibroblast transition through both the WNT/β-Catenin and Hedgehog/GLI signaling pathways in vitro. A mRNA expression of endogenous SENP1 , β-Catenin, and GLI1 were assessed using RT-PCR in different groups. RT-PCR analysis indicating that β-Catenin and GLI1 mRNA levels did not change in the presence of LV - SENP1-shRNA during TGFβ1 induction. *P < 0.01, **P < 0.05, ▲ P < 0.01, ▲▲ P > 0.05. *, the control group vs . the TGFβ1 group; **, the SENP1 knockdown group vs. the TGFβ1 group; ▲ , the control group vs. the TGFβ1 group; ▲▲ , the SENP1 knockdown group vs. the TGFβ1 group. B SENP1 level in total cell lysates was assessed using western blot analyses. *P < 0.01, **P < 0.05. *, the control group vs. the TGFβ1 group; **, the SENP1 knockdown group vs. the TGFβ1 group. C β-Catenin and GLI1 levels in total cell lysates were assessed using western blot analyses. The SUMOylation analysis of the immunoprecipitated β-Catenin and GLI1 proteins. β-Catenin and GLI1 were immunoprecipitated from whole-cell lysates using anti-β-Catenin and anti-GLI1 antibodies, respectively. The blots were probed with SUMO1. *P < 0.01, **P < 0.05, ▲ P < 0.01, ▲▲ P < 0.05. *, the control group vs . the TGFβ1 group; ** , the SENP1 knockdown group vs. the TGFβ1 group (β-Catenin and GLI1); ▲ , the control group vs . the TGFβ1 group; ▲▲ , the SENP1 knockdown group vs. the TGFβ1 group (β-Catenin-SUMO and GLI1-SUMO). All data are presented as the mean ± SEM of three independent experiments. Quantification is shown in the lower panel. Each experiment was performed in triplicate.

Article Snippet: Recombinant TGFβ1 (Proteintech Group, Wuhan, China) was used at a 10 ng/mL concentration to induce LR-MSC myofibroblast transition.

Techniques: Protein-Protein interactions, In Vitro, Expressing, Reverse Transcription Polymerase Chain Reaction, shRNA, Control, Knockdown, Western Blot, Immunoprecipitation

Journal: Scientific Reports

Article Title: TGFβ1 secreted by cancer-associated fibroblasts induces epithelial-mesenchymal transition of bladder cancer cells through lncRNA-ZEB2NAT

doi: 10.1038/srep11924

Figure Lengend Snippet: A : TGFβ1 in conditional mediums secreted by 5637 (CTRL), NF and CAF cells were quantified by ELISA. B : The mRNA expression levels of TGFβ1 in CAFs, NFs and three bladder cancer cell lines by qRT-PCR using β-actin gene as the normalization control. C : The expression of phosphorylated Smad2 and total Smad2 protein in the CAF-CM treated bladder cancer cell lines by immunoblotting. β-actin protein was used as the loading control. D : The expression levels of TGFβ1 and TGFβRII in bladder cancer cell lines cultured with CAF-CM were detected by qRT-PCR using β-actin gene as the normalization control. *** P < 0.001.

Article Snippet: The concentrations of TGFβ1 in different media were measured using human TGFβ1 ELISA kit (BOSTER, EK0513, Wuhan, China), according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Control, Western Blot, Cell Culture

Journal: Scientific Reports

Article Title: TGFβ1 secreted by cancer-associated fibroblasts induces epithelial-mesenchymal transition of bladder cancer cells through lncRNA-ZEB2NAT

doi: 10.1038/srep11924

Figure Lengend Snippet: A : Summary of altered expression of lncRNAs in the CAF-CM-treated 5637 and J82 cells, comparing to the NF-CM-treated ones. 1.5-fold of relative mRNA level using qRT-PCR, normalized by β-actin gene, was used as the threshold for significant changes. B and C : ZEB2NAT expression levels in the CAF-CM treated 5637 and J82 cells upon the treatments of a TGFβ1 neutralizing antibody ( B ) and a TGFβRI inhibitor (SB-431542) ( C ), respectively. D : Exogenous expression of ZEB2NAT lncRNA in 5637 cells, detected by qRT-PCR using β-actin gene as the normalization control. E – G : Effects of ZEB2NAT lncRNAs overexpression on cell invasion by the Transwell invasion assay ( E , F ) and protein levels of E-cadherin, ZEB1 and ZEB2 by immunoblotting ( G ) in 5637 cells. H: Knockdown of ZEB2NAT by RNAi using two siRNAs targeting different regions of the lncRNA (siZEB2NAT-1 and siZEB2NAT-2) in 5637 cells. The relative ZEB2NAT expression levels were at 48 hours after siRNA transfection and determined by qRT-PCR using β-actin gene as the normalization control. I , J , K : Effects of ZEB2NAT lncRNAs knockdown on cell invasion by the Transwell invasion assay ( I , J ) and protein levels of E-cadherin and ZEB2 by immunoblotting ( K ) in the CAF-CM treated 5637 cells. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The concentrations of TGFβ1 in different media were measured using human TGFβ1 ELISA kit (BOSTER, EK0513, Wuhan, China), according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Control, Over Expression, Transwell Invasion Assay, Western Blot, Knockdown, Transfection

Journal: Scientific Reports

Article Title: TGFβ1 secreted by cancer-associated fibroblasts induces epithelial-mesenchymal transition of bladder cancer cells through lncRNA-ZEB2NAT

doi: 10.1038/srep11924

Figure Lengend Snippet: The expression levels of ZEB2NAT ( A ) and TGFβ1 transcripts ( B ), and ZEB2 protein ( C ) in 30 human bladder cancer tissues (Tumor or T) and the paired normal tissues (Normal or N) from the same patient. Relative ZEB2NAT and TGFβ1 mRNA levels were assessed by qRT-PCR and normalized by β-actin gene. D – F : The correlations among ZEB2NAT, TGFβ1 and ZEB2 were decided by Pearson correlation analysis. ΔCT values in qRT-PCR were used as the expression levels of ZEB2NAT and TGFβ1 transcripts. ZEB2 protein bands on Western blot were quantified by Image J software. P < 0.05 was considered statistically significant.

Article Snippet: The concentrations of TGFβ1 in different media were measured using human TGFβ1 ELISA kit (BOSTER, EK0513, Wuhan, China), according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software

Journal: iScience

Article Title: CD168 + macrophages promote hepatocellular carcinoma tumor stemness and progression through TOP2A /β-catenin/ YAP1 axis

doi: 10.1016/j.isci.2023.106862

Figure Lengend Snippet:

Article Snippet: Human TGF-beta1 ELISA Kit , proteintech , Cat # KE00002.

Techniques: Recombinant, Preserving, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, SYBR Green Assay, Software