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: 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