Journal: Cancer Medicine

Article Title: The Biological and Prognostic Implications of the Nicotinic Acetylcholine Receptor α 3, α 5, and α 7 Subunits in Oral Squamous Cell Carcinoma

doi: 10.1002/cam4.71358

Figure Lengend Snippet: Influences of CHRNA3 , CHRNA5 , and CHRNA7 expression in SCC‐4 tongue cancer cells. (A) The effects of nicotine (1 μM) treatment (qRT‐PCR). (B1–B3) The effects of overexpressed CHRNA3 , CHRNA5 , and CHRNA7 , respectively (qRT‐PCR). (C1–C3) The effects of overexpressed DSG2 , TGF β 1 treatment (2.5 ng/mL), and overexpressed TGFBR2 , respectively (qRT‐PCR). (D1–D2) Summary of the qRT‐PCR results. (E) The expression patterns of our SCC‐4 cells at two different time points (SCC‐4‐1 and SCC‐4‐2) and the DepMap SCC‐4 cells (RNA‐seq). (F1–F2) GSEA results comparing SCC‐4‐2 cells to SCC‐4‐1 cells. qRT‐PCR, quantitative reverse transcription polymerase chain reaction. RNA‐seq, RNA sequencing. TPM, transcripts per million. GO, gene ontology. BP, biological process. CC, cellular component. *, p < 0.05. **, p < 0.01. ***, p < 0.001.

Article Snippet: Nicotine (1 μM) (TargetMol) and TGF β 1 (2.5 ng/mL) (Gibco) were used as stimulants for nAChRs and TGFBRs, respectively.

Techniques: Expressing, Quantitative RT-PCR, RNA Sequencing, Reverse Transcription, Polymerase Chain Reaction

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: 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: 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: Journal of Inflammation (London, England)

Article Title: Protective effects of thalidomide on pulmonary injuries in a rat model of paraquat intoxication

doi: 10.1186/s12950-015-0093-0

Figure Lengend Snippet: Effects of thalidomide on serum levels of IL-6, TNF-α, TGF-β1 and COL1A1 induced by PQ. Blood samples from rats from each treatment group were collected through retro-orbital bleeding at days 1, 2, 3, 5, 7, 10 and 15 after paraquat exposure and serum levels of IL-6 ( a ), TNF-α ( b ), TGF-β1 ( c ) and COL1A1 ( d ) were measured by ELISA. The values represent mean ± SD from each group (● Control, ■ PQ group and ▲ PQ + Thal group). * p <0.05 compared to Control, # p <0.05 compared to Paraquat

Article Snippet: Paraquat was provided by Shandong Yinuo Company (Shandong, China); Thalidomide was purchased from Changzhou Pharmaceutical Company (Changzhou, China); Polyclonal antibodies against TNF-α, IL-6, TGFβ1 and collagen-1 were purchased from Boster Biological Technology (Wuhan, China); Horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody and streptavidin-peroxidase immunohistochemistry kit were provided by Beijing Zhongshan-Golden Bridge Biological Technology (Beijing, China); Trizol and SuperScript II Reverse Transcriptase were obtained from Invitrogen (Carlsbad, USA); Taq DNA polymerase and DNA size marker were purchased from Beijing Dingguo Changsheng Biotechnology (Beijing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Inflammation (London, England)

Article Title: Protective effects of thalidomide on pulmonary injuries in a rat model of paraquat intoxication

doi: 10.1186/s12950-015-0093-0

Figure Lengend Snippet: Effects of thalidomide on expression levels of IL-6, TNF-α, TGF-β1 and COL1A1 in lung tissues after PQ intoxication. Lung tissues from each treatment group were excised on day 15 after PQ administration and mRNA expression levels of IL-6, TNF-α, TGF-β1 and COL1A1 genes were examined using RT-PCR

Article Snippet: Paraquat was provided by Shandong Yinuo Company (Shandong, China); Thalidomide was purchased from Changzhou Pharmaceutical Company (Changzhou, China); Polyclonal antibodies against TNF-α, IL-6, TGFβ1 and collagen-1 were purchased from Boster Biological Technology (Wuhan, China); Horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody and streptavidin-peroxidase immunohistochemistry kit were provided by Beijing Zhongshan-Golden Bridge Biological Technology (Beijing, China); Trizol and SuperScript II Reverse Transcriptase were obtained from Invitrogen (Carlsbad, USA); Taq DNA polymerase and DNA size marker were purchased from Beijing Dingguo Changsheng Biotechnology (Beijing, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Inflammation (London, England)

Article Title: Protective effects of thalidomide on pulmonary injuries in a rat model of paraquat intoxication

doi: 10.1186/s12950-015-0093-0

Figure Lengend Snippet: Immunohistochemistry detection of the expression of IL-6, TNF-α, TGF-β1 and COL1A1 proteins in lung tissues after PQ intoxication. Lung tissues from Control ( a , d , g , j ), PQ ( b , e , h , k ) and PQ + Thal ( c , f , i , l ) groups were collected on day 15 after PQ administration and expression of IL-6 ( a , b , c ), TNF-α ( d , e , f ), TGF-β1 ( g , h , i ) and COL1A1 ( j , k , l ) in lung tissues were examined using immunohistochemical staining (×100)

Article Snippet: Paraquat was provided by Shandong Yinuo Company (Shandong, China); Thalidomide was purchased from Changzhou Pharmaceutical Company (Changzhou, China); Polyclonal antibodies against TNF-α, IL-6, TGFβ1 and collagen-1 were purchased from Boster Biological Technology (Wuhan, China); Horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody and streptavidin-peroxidase immunohistochemistry kit were provided by Beijing Zhongshan-Golden Bridge Biological Technology (Beijing, China); Trizol and SuperScript II Reverse Transcriptase were obtained from Invitrogen (Carlsbad, USA); Taq DNA polymerase and DNA size marker were purchased from Beijing Dingguo Changsheng Biotechnology (Beijing, China).

Techniques: Immunohistochemistry, Expressing, Control, Immunohistochemical staining, Staining