Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: YAP1 is upregulated in HBZY-1 cells treated with high glucose. (A) RT-qPCR was used to measure the expression of YAP1 in HBZY-1 cells treated with 30 mmol/l serum-free DMEM medium for 0, 6, 12, 24 and 48 h, respectively. (B) Western blotting was used to measure the expression of YAP1 in HBZY-1 cells treated with 30 mmol/l serum-free DMEM medium for 0, 6, 12, 24 and 48 h, respectively. (C) Transfection efficiency of YAP1 small interfering RNA was detected by RT-qPCR. *** P<0.001 vs. HG 0 h; ### P<0.001 vs. HG + si-NC. YAP1, Yes-associated protein 1; RT-qPCR, reverse transcription-quantitative PCR; si-, small interfering RNA; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Transfection, Small Interfering RNA, Real-time Polymerase Chain Reaction, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: Knockdown of YAP1 inhibited the proliferation of HBZY-1 cells induced by high glucose. (A) Effect of YAP1 knockdown on the viability of HBZY-1 cells treated with high glucose was detected by Cell Counting Kit-8 assay. (B) Effect of YAP1 knockdown on the expression of PCNA in the HBZY-1 cells treated with high glucose was measured by western blotting. * P<0.05 vs. HG + si-NC. YAP1, Yes-associated protein 1; PCNA, proliferating cell nuclear antigen; si-, small interfering RNA; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Cell Counting, Expressing, Western Blot, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: Knockdown of YAP1 suppresses the inflammation of HBZY-1 cells induced by high glucose. (A) Effect of YAP1 knockdown on the expressions of pro-inflammatory factors (TNF-α, IL-1β and IL-6) was detected by enzyme-linked immunosorbent assay. (B) Effect of YAP1 knockdown on the expression of MCP-1 was detected by reverse transcription-quantitative PCR. (C) Effect of YAP1 knockdown on the expression of MCP-1 was detected by western blotting. ** P<0.01 and *** P<0.001 vs. HG + si-NC. YAP1, Yes-associated protein 1; si-, small interfering RNA; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol; TNF, tumor necrosis factor; IL, interleukin; MCP-1, monocyte chemoattractant protein-1.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: Knockdown of YAP1 inhibits the deposition of extracellular matrix and fibrosis in HBZY-1 cells induced by high glucose. (A) Effect of YAP1 knockdown on the expressions of collagen IV, α-SMA and FN was detected by western blotting. (B and C) Effect of YAP1 knockdown on the expressions of collagen IV was detected by immunofluorescence. Scale bars, 50 µm. (D) Effect of YAP1 knockdown on the expressions of fibrogenic factors CTGF and TGF-β1 was detected by western blotting. * P<0.05, ** P<0.01 and *** P<0.001 vs. HG + si-NC. YAP1, Yes-associated protein 1; si-, small interfering RNA; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol; SMA, smooth muscle actin; FN, fibronectin; CTGF, connective tissue growth factor; TGF, transforming growth factor.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Western Blot, Immunofluorescence, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: Knockdown of YAP1 inhibited NF-κB/JMJD3 signal pathway in HBZY-1 cells induced by high glucose. (A) Effect of YAP1 knockdown on the phosphorylation levels of IκBα and p65 were detected by western blotting. (B) Effect of YAP1 knockdown on the expression of JMJD3 was detected by western blotting. * P<0.05, ** P<0.01 and *** P<0.001 vs. HG + si-NC. YAP1, Yes-associated protein 1; si-, small interfering RNA; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol; NF-κB, nuclear factor-κB; JMJD3, Jumonji domain-containing protein D3; p-, phosphorylated-; IκBα, inhibitor of NF-κB.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Western Blot, Expressing, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: Inhibitors of NF-κB/JMJD3 signal pathway attenuates the facilitatory effects of YAP1 on the proliferation and inflammation of HBZY-1 cells induced by high glucose. (A) The efficiency of YAP1 overexpression was determined by RT-qPCR. (B) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression levels of IκBα and p65 were detected by western blotting. (C) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the viability of HBZY-1 cells treated with high glucose was detected by Cell Counting Kit-8 assay. (D) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression of PCNA in the HBZY-1 cells treated with high glucose was measured by western blotting. (E) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expressions of pro-inflammatory factors (TNF-α, IL-1β and IL-6) was detected by enzyme-linked immunosorbent assay. (F) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression of MCP-1 was detected by RT-qPCR. (G) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression of MCP-1 was detected by western blotting. ** P<0.01 vs. HG + Oe-NC. # P<0.05, ## P<0.01 and ### P<0.001 vs. HG + Oe-YAP1. YAP1, Yes-associated protein 1; RT-qPCR, reverse transcription-quantitative PCR; Oe, overexpression vector; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol; NF-κB, nuclear factor-κB; JMJD3, Jumonji domain-containing protein D3; p-, phosphorylated-; IκBα, inhibitor of NF-κB; TNF, tumor necrosis factor; IL, interleukin; MCP-1, monocyte chemoattractant protein-1.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Western Blot, Cell Counting, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Plasmid Preparation, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: YAP1 promotes high glucose-induced inflammation and extracellular matrix deposition in glomerular mesangial cells by modulating NF-κB/JMJD3 pathway

doi: 10.3892/etm.2021.10784

Figure Lengend Snippet: Inhibitors of NF-κB/JMJD3 signal pathway attenuates the facilitatory effects of YAP1 on the deposition of extracellular matrix and fibrosis of HBZY-1 cells induced by high glucose. (A) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression levels of collagen IV, α-SMA and FN was detected by western blotting. (B and C) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression levels of collagen IV was detected by immunofluorescence. Scale bars, 50 µm. (D) Effect of the inhibitors of NF-κB/JMJD3 signal pathway on the expression levels of fibrogenic factors CTGF and TGF-β1 was detected by western blotting. # P<0.05 and ## P<0.01 vs. HG + Oe-YAP1. YAP1, Yes-associated protein 1; Oe, overexpression vector; NC, negative control; HG, high-glucose group; NG, normal control group; MA, mannitol; NF-κB, nuclear factor-κB; JMJD3, Jumonji domain-containing protein D3; SMA, smooth muscle actin; FN, fibronectin; CTGF, connective tissue growth factor; TGF, transforming growth factor.

Article Snippet: The YAP1 overexpression vector pcDNA-TRIM31 (Oe-YAP1) and empty control vector pcDNA (Oe-NC) were constructed by Shanghai GenePharma Co., Ltd. Small interfering RNA (siRNA) against YAP1 (si-YAP1) and siRNA negative control (si-NC) were synthesized by Shanghai GenePharma Co., Ltd. A final concentration of 100 nM plasmids were transfected into cells using Lipofectamine ® 2000 (Thermo Fisher Scientific, Inc.) transfection reagent.

Techniques: Expressing, Western Blot, Immunofluorescence, Over Expression, Plasmid Preparation, Negative Control