Journal: Oncotarget

Article Title: Induction of methionine adenosyltransferase 2A in tamoxifen-resistant breast cancer cells

doi: 10.18632/oncotarget.5298

Figure Lengend Snippet: A. Immunoblot analysis of MAT2 in MCF-7 and TAMR-MCF-7 cells. Each lane represents different sample. B. Basal MAT2A promoter reporter activities in MCF-7 and TAMR-MCF-7 cells. MCF-7 and TAMR-MCF-7 cells were seeded in 12 wells plate for 1 day. Both the cell types (60% confluency) were then transiently co-transfected with MAT2A-luc reporter plasmid containing −570/+61 bp human MAT2A promoter (1 μg/well) and phRL-SV (hRenilla) (1 ng/well). Dual luciferase reporter assays were performed on the lysed cells 18 h after transfection in serum free condition. Reporter gene activity was calculated as a relative ratio of firefly luciferase to hRenilla luciferase activity. Data represent mean ± SD with 6 different samples (significant versus MCF-7 cells, ** P < 0.01). C. MAT2A mRNA levels in MCF-7 and TAMR-MCF-7 cells. MAT2A mRNA levels were determined by quantitative RT-PCR. Data represent the mean ± SD ( n = 4)(significant versus MCF-7 cells, ** P < 0.01). D. Immunohistochemistry of MAT2A in human breast cancer tissues. Four TAM-responsive and four TAM-resistant cases were estimated. The brown color staining represents MAT2A expression. When we determined immunoreactivity in IgG-incubated breast cancer tissue samples (negative control), we could not detect any positive staining. E. MAT2A immunoblot analyses in T47D cells. The basal MAT2A levels were compared in T47D, MCF-7, TAMR-MCF-7 and MDA-MB-231 cells.

Article Snippet: Sections were then incubated overnight with anti-MAT2A antibody (NBP1-28605; Novus biological, CO, USA) at 4°C.

Techniques: Western Blot, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Quantitative RT-PCR, Immunohistochemistry, Staining, Expressing, Incubation, Negative Control

Journal: Oncotarget

Article Title: Induction of methionine adenosyltransferase 2A in tamoxifen-resistant breast cancer cells

doi: 10.18632/oncotarget.5298

Figure Lengend Snippet: A. AP-1 activities in MCF-7 and TAMR-MCF-7 cells. Left; AP-1 minimal reporter activity. MCF-7 and TAMR-MCF-7 cells were co-transfected with pAP-1-Luc reporter (1 μg/well) and phRL-SV plasmids (hRenilla, 1 ng/well). Luciferase reporter activity was determined described as figure legend of Figure . Data represent mean ± SD with 6 different samples (significant versus MCF-7 cells, ** P < 0.01). Right; Nuclear levels of c-Jun, c-Fos, Jun-B and Jun-D in MCF-7 and TAMR-MCF-7 cells. Each AP-1 protein was detected in the nuclear fractions isolated from serum-deprived MCF-7 and TAMR-MCF-7 cells. B. Left; Effect of c-Jun siRNA on MAT2A expression level in TAMR-MCF-7 cells. TAMR-MCF-7 cells were transfected with control or c-Jun siRNA (60 p mole/well) for 36 h. The protein levels of MAT2 and c-Jun in total cell lysates were determined by immunoblotting. Middle; Effect of c-Jun siRNA on MAT2A gene transcription in MCF-7 and TAMR-MCF-7 cells. Both cell lines were co-transfected with MAT2A-luc reporter plasmid containing −570/+61 bp human MAT2A promoter (1 μg/well), phRL-SV (1 ng/well) and control or c-Jun siRNA (20 p mole/well) for 36 h. Luciferase reporter activity was determined described as figure legend of figure . Data represent mean ± SD with 3 different samples. Right; Effect of c-Jun siRNA on AP-1 reporter activity in TAMR-MCF-7 cells. TAMR-MCF-7 cells were co-transfected with control or c-Jun siRNA (20 p mole/well) and pAP-1-Luc (1 μg/well)/phRL-SV (1 ng/well) for 36 h. Data represent mean ± SD with 6 different samples (significant versus control siRNA-transfected TAMR-MCF-7 cells, ** P < 0.01). C. Nrf2/ARE activities in MCF-7 and TAMR-MCF-7 cells. Upper; nuclear level of Nrf2 were detected in the nuclear fractions isolated from serum-deprived MCF-7 and TAMR-MCF-7 cells. Lower; the basal ARE reporter activities in MCF-7 and TAMR-MCF-7 cells. Both cell lines were transiently co-transfected with pGL-ARE-luc plasmid (1 μg/well) and phRL-SV (1 ng/well). Dual luciferase reporter assays were performed on the lysed cells 18 h after transfection in serum free condition. Data represent mean ± SD with 6 different samples (significant versus MCF-7 cells, ** P < 0.01). D. Effect of Nrf2 siRNA on MAT2A expression (upper) and MAT2A gene transcription (lower) in TAMR-MCF-7 cells. Upper; MCF-7 cell and TAMR-MCF-7 cells were transfected with control or Nrf2 siRNA (60 p mole/well) for 36 h. MAT2 and Nrf2 protein expression was determined by immunoblotting. Lower; effect of Nrf2 siRNA on MAT2A gene transcription in MCF-7 and TAMR-MCF-7 cells. Both cell lines were co-transfected with MAT2A-luc reporter plasmid containing −570/+61 bp human MAT2A promoter (1 μg/well), phRL-SV (1 ng/well) and control or Nrf2 siRNA (20 p mole/well) for 36 h. Data represent mean ± SD with 3 different samples.

Article Snippet: Sections were then incubated overnight with anti-MAT2A antibody (NBP1-28605; Novus biological, CO, USA) at 4°C.

Techniques: Activity Assay, Transfection, Luciferase, Isolation, Expressing, Control, Western Blot, Plasmid Preparation

Journal: Oncotarget

Article Title: Induction of methionine adenosyltransferase 2A in tamoxifen-resistant breast cancer cells

doi: 10.18632/oncotarget.5298

Figure Lengend Snippet: A. NF-κB activation in TAMR-MCF-7 cells. Upper; Nuclear level of p65. p65 protein levels were detected in the nuclear fractions isolated from serum-deprived MCF-7 and TAMR-MCF-7 cells. Lower; NF-κB minimal reporter activity. MCF-7 and TAMR-MCF-7 cells in 12 well plates were co-transfected with NF-κB-luc reporter (1 μg/well) and phRL-SV plasmids (1 ng/well) for 18 h in serum free condition. Luciferase reporter activity was determined described as figure legend of Figure . Data represent mean ± SD with 6 different samples (significant versus MCF-7 cells, ** P < 0.01). B. Effects of TPCK (NF-κB inhibitor) on MAT2A expression (upper) and NF-κB reporter activity (lower) in TAMR-MCF-7 cells. Upper; TAMR-MCF-7 cells were exposed to TPCK with the indicated concentration for 24 h. MAT2 expression level was determined from total cell lysates using immunoblotting. Lower; MCF-7 and TAMR-MCF-7 cells were co-transfected with NF-κB-luc reporter (1 μg/well) and phRL-SV plasmids (1 ng/well). Concomitantly, TPCK with the indicated concentration was treated to the transfected TAMR-MCF-7 cells for 24 h. Luciferase reporter activity was determined described as figure legend of figure . Data represent mean ± SD with 3 different samples (significant versus MCF-7 cells, ** P < 0.01; significant versus vehicle-treated TAMR-MCF-7 cells, ## P < 0.01). C. Effect of TPCK on MAT2A gene transcription. MCF-7 and TAMR-MCF-7 cells were co-transfected with MAT2A-luc reporter plasmid (1 μg/well) and phRL-SV plasmids (1 ng/well). Concomitantly, TPCK with the indicated concentration was treated to the transfected TAMR-MCF-7 cells for 24 h. Data represent mean ± SD with 3 different samples (significant versus MCF-7 cells, ** P < 0.01; significant versus vehicle-treated TAMR-MCF-7 cells, # P < 0.05). D. Effect of IκBα overexpression on the nuclear expression of p65 and MAT2A gene transcription in TAMR-MCF-7 cells. TAMR-MCF-7 cells were co-transfected with MAT2A-luc reporter plasmid (1 μg/well) and pCMV5 or IκBα overexpression plasmid (0.5 μg, respectively), and phRL-SV plasmids (1 ng/well) for 18 h in serum free condition. Data represent mean ± SD with 4 different samples (significant versus pCMV5-transfected MCF-7 cells, ** P < 0.01; significant versus pCMV5-transfected TAMR-MCF-7 cells, # P < 0.05).

Article Snippet: Sections were then incubated overnight with anti-MAT2A antibody (NBP1-28605; Novus biological, CO, USA) at 4°C.

Techniques: Activation Assay, Isolation, Activity Assay, Transfection, Luciferase, Expressing, Concentration Assay, Western Blot, Plasmid Preparation, Over Expression

Journal: Oncotarget

Article Title: Induction of methionine adenosyltransferase 2A in tamoxifen-resistant breast cancer cells

doi: 10.18632/oncotarget.5298

Figure Lengend Snippet: A. Down-regulation of miR-146a and miR-146b expression in TAMR-MCF-7 cells. miR-146a and miR-146b expression in MCF-7 and TAMR-MCF-7 cells were determined by using miScript PCR kit with miScript primers specific for mature miR-146a and miR-146b. Samples were normalized to small nRNA U6. B. Effects of miR-146a and miR-146b mimics on the miR-146a/b expression in TAMR-MCF-7 cells. TAMR-MCF-7 cells were cultured in 6 wells plate and then transfected with miR-146a or miR-146b mimics for 36 h (120 p mole/well). miR-146a and miR-146b levels were determined by using miScript PCR kit with miScript primers specific for mature miR-146a and miR-146b. Samples were normalized to small nRNA U6. C. Effects of miR-146a/b mimic on NF-κB activity and MAT2A protein expression in TAMR-MCF-7 cells. TAMR-MCF-7 cells were cultured in 6 wells plate and then transfected with miR-146a or miR-146b mimics (120 p mole/well) for 36 h and immunoblottings were performed. The data were confirmed by two independent experiments. D. Effects of miR-146b mimic on MAT2A mRNA expression in TAMR-MCF-7 cells. TAMR-MCF-7 cells were cultured in 6 wells plate and then transfected with miR-146a or miR-146b mimics (120 p mole/well) for 36 h. MAT2A mRNA levels were determined by quantitative RT-PCR. Data represent mean ± SD with 3 different samples (significant versus control mimic miR-treated TAMR-MCF-7 cells, * P < 0.05). E. miR-146b expression in T47D cells. miR-146b expression in MCF-7 and T47D cells were determined by using miScript PCR kit with miScript primers specific for mature miR-146b. Samples were normalized to small nRNA U6.

Article Snippet: Sections were then incubated overnight with anti-MAT2A antibody (NBP1-28605; Novus biological, CO, USA) at 4°C.

Techniques: Expressing, Cell Culture, Transfection, Activity Assay, Quantitative RT-PCR, Control

Journal: Oncotarget

Article Title: Induction of methionine adenosyltransferase 2A in tamoxifen-resistant breast cancer cells

doi: 10.18632/oncotarget.5298

Figure Lengend Snippet: A. Effect of MAPK inhibitors and PI3K inhibitor on MAT2A expression. Upper; MAT2A protein expression was completely suppressed by 24 h incubation of LY294002 (LY, 20 μM), a PI3K inhibitor in TAMR-MCF-7 cells. PD98059 (PD, 20 μM, ERK inhibitor), SB203580 (SB, 10 μM, p38 kinase inhibitor) and SP600125 (SP, 10 μM, JNK inhibitor) were also used. Lower; Densitometry data. Data represent mean ± SD with 3 different samples (significant versus control MCF-7 cells, ** P < 0.01; significant versus TAMR-MCF-7 cells, ## P < 0.01). B. Effect of Mycp85 (dominant negative mutant form of PI3K) overexpression on MAT2A gene transcription. TAMR-MCF-7 cells were co-transfected with MAT2A-luc reporter plasmid (1 μg/well) phRL-SV plasmids (1 ng/well) and pCMV5 or Mycp85 overexpression plasmid (0.5 μg/well) for 18 h in serum free condition. Luciferase reporter activity was determined described as figure legend of Figure . Data represent mean ± SD with 3 different samples (significant versus pCMV5-transfected MCF-7 cells, ** P < 0.01; significant versus pCMV5-transfected TAMR-MCF-7 cells, ## P < 0.01). C. Effect of PI3K inhibition on NF-κB activity. Left; nuclear p65, nuclear Nrf2, and nuclear c-Jun detection. TAMR-MCF-7 cells were treated with LY294002 (LY, 20 μM) for 24 h and nuclear levels of p65, Nrf2, and c-Jun were detected by immunoblotting. Right; Effect of Mycp85 overexpression on NF-κB minimal reporter activity. TAMR-MCF-7 cells were co-transfected with pNF-κB-Luc reporter plasmid (1 μg/well) and pCMV5 or Mycp85 overexpression plasmid (0.5 μg/well). Then, pNF-κB-Luc reporter activity was determined 18 h after transfection in serum-free condition. Data represent mean ± SD with 3 different samples (significant versus pCMV5-transfected MCF-7 cells, ** P < 0.01; significant versus pCMV5-transfected TAMR-MCF-7 cells, ## P < 0.01). D. Effect of LY294002 (PI3K inhibitor, LY, 20 μM) on miR-146b expression in TAMR-MCF-7 cells. TAMR-MCF-7 cells was exposed to LY294002 for 24 h and then miR-146b level was determined by using miScript PCR. Samples were normalized to small nRNA U6. Data represent mean ± SD with 3 different samples (significant versus vehicle-treated TAMR-MCF-7 cells, ** P < 0.01).

Article Snippet: Sections were then incubated overnight with anti-MAT2A antibody (NBP1-28605; Novus biological, CO, USA) at 4°C.

Techniques: Expressing, Incubation, Control, Dominant Negative Mutation, Over Expression, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Inhibition, Western Blot

Journal: Oncotarget

Article Title: Induction of methionine adenosyltransferase 2A in tamoxifen-resistant breast cancer cells

doi: 10.18632/oncotarget.5298

Figure Lengend Snippet: The scheme shows positive feedback loop between the PTEN-controlled PI3K/Akt pathway and miR-146b-controlled NF-κB/MAT2A expression.

Article Snippet: Sections were then incubated overnight with anti-MAT2A antibody (NBP1-28605; Novus biological, CO, USA) at 4°C.

Techniques: Expressing

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: Primer sequences used for real-time PCR.

Article Snippet: The cells were seeded on six-well plates overnight, pretreated with the MAT2A inhibitor PF9366 (S0435; Selleck Chemicals, USA), SAM (S5109; Selleck Chemicals, USA), or small interference RNA transfection if necessary, followed by incubation with antibiotic-free DMEM and 10% FBS, and at a density of 2 × 10 5 cells/2 mL per well before infection.

Techniques: Sequencing

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: MAT2A expression was upregulated in inflamed gingival tissues.

Article Snippet: The cells were seeded on six-well plates overnight, pretreated with the MAT2A inhibitor PF9366 (S0435; Selleck Chemicals, USA), SAM (S5109; Selleck Chemicals, USA), or small interference RNA transfection if necessary, followed by incubation with antibiotic-free DMEM and 10% FBS, and at a density of 2 × 10 5 cells/2 mL per well before infection.

Techniques: Expressing

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: MAT2A inhibition decreased P. gingivalis -induced inflammatory responses in hGfs.

Article Snippet: The cells were seeded on six-well plates overnight, pretreated with the MAT2A inhibitor PF9366 (S0435; Selleck Chemicals, USA), SAM (S5109; Selleck Chemicals, USA), or small interference RNA transfection if necessary, followed by incubation with antibiotic-free DMEM and 10% FBS, and at a density of 2 × 10 5 cells/2 mL per well before infection.

Techniques: Inhibition

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: MAT2A-knockdown reduced inflammation in P. gingivalis -infected hGFs.

Article Snippet: The cells were seeded on six-well plates overnight, pretreated with the MAT2A inhibitor PF9366 (S0435; Selleck Chemicals, USA), SAM (S5109; Selleck Chemicals, USA), or small interference RNA transfection if necessary, followed by incubation with antibiotic-free DMEM and 10% FBS, and at a density of 2 × 10 5 cells/2 mL per well before infection.

Techniques: Knockdown, Infection

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: NF-κB/MAPK pathway was involved in the enhanced MAT2A expression.

Article Snippet: The cells were seeded on six-well plates overnight, pretreated with the MAT2A inhibitor PF9366 (S0435; Selleck Chemicals, USA), SAM (S5109; Selleck Chemicals, USA), or small interference RNA transfection if necessary, followed by incubation with antibiotic-free DMEM and 10% FBS, and at a density of 2 × 10 5 cells/2 mL per well before infection.

Techniques: Expressing

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: Primer sequences used for real-time PCR.

Article Snippet: Proteins at 10 μL per lane were separated by 4–20% SDS-PAGE (Beyotime, China) and then transferred onto PVDF membranes (Merck, Germany), which were blocked with 5% bovine albumin and then incubated with primary antibodies to MAT2A (1:1000; 55309–1-AP; Proteintech, China), IL-1β (1:1000; BJ11258993; Bioss, China), TNF-α (1:400; GB11188; Servicebio, China), IL-6 (1:1000; 21865–1-AP; Proteintech, China), MCP-1 (1:1500; ab25124; Abcam, USA), P65 (1:1000; 8242S; CST, Germany), Erk (1:1500; GB11560; Servicebio, China), P38 (1:1000;8690S; CST, Germany), JNK (1:1000; AF6318; Affinity, USA), p-P65 (1:1000; 3033S; CST, Germany), p-Erk (1:1000; AF1015; Affinity, USA), p-P38 (1:1000; 4511S; CST, Germany), p-JNK (1:1000;4668S; CST, Germany) and β-ACTIN (1:50000; 66009-l-Ig; Proteintech, China).

Techniques: Sequencing

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: MAT2A expression was upregulated in inflamed gingival tissues.

Article Snippet: Proteins at 10 μL per lane were separated by 4–20% SDS-PAGE (Beyotime, China) and then transferred onto PVDF membranes (Merck, Germany), which were blocked with 5% bovine albumin and then incubated with primary antibodies to MAT2A (1:1000; 55309–1-AP; Proteintech, China), IL-1β (1:1000; BJ11258993; Bioss, China), TNF-α (1:400; GB11188; Servicebio, China), IL-6 (1:1000; 21865–1-AP; Proteintech, China), MCP-1 (1:1500; ab25124; Abcam, USA), P65 (1:1000; 8242S; CST, Germany), Erk (1:1500; GB11560; Servicebio, China), P38 (1:1000;8690S; CST, Germany), JNK (1:1000; AF6318; Affinity, USA), p-P65 (1:1000; 3033S; CST, Germany), p-Erk (1:1000; AF1015; Affinity, USA), p-P38 (1:1000; 4511S; CST, Germany), p-JNK (1:1000;4668S; CST, Germany) and β-ACTIN (1:50000; 66009-l-Ig; Proteintech, China).

Techniques: Expressing

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: MAT2A inhibition decreased P. gingivalis -induced inflammatory responses in hGfs.

Article Snippet: Proteins at 10 μL per lane were separated by 4–20% SDS-PAGE (Beyotime, China) and then transferred onto PVDF membranes (Merck, Germany), which were blocked with 5% bovine albumin and then incubated with primary antibodies to MAT2A (1:1000; 55309–1-AP; Proteintech, China), IL-1β (1:1000; BJ11258993; Bioss, China), TNF-α (1:400; GB11188; Servicebio, China), IL-6 (1:1000; 21865–1-AP; Proteintech, China), MCP-1 (1:1500; ab25124; Abcam, USA), P65 (1:1000; 8242S; CST, Germany), Erk (1:1500; GB11560; Servicebio, China), P38 (1:1000;8690S; CST, Germany), JNK (1:1000; AF6318; Affinity, USA), p-P65 (1:1000; 3033S; CST, Germany), p-Erk (1:1000; AF1015; Affinity, USA), p-P38 (1:1000; 4511S; CST, Germany), p-JNK (1:1000;4668S; CST, Germany) and β-ACTIN (1:50000; 66009-l-Ig; Proteintech, China).

Techniques: Inhibition

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: MAT2A-knockdown reduced inflammation in P. gingivalis -infected hGFs.

Article Snippet: Proteins at 10 μL per lane were separated by 4–20% SDS-PAGE (Beyotime, China) and then transferred onto PVDF membranes (Merck, Germany), which were blocked with 5% bovine albumin and then incubated with primary antibodies to MAT2A (1:1000; 55309–1-AP; Proteintech, China), IL-1β (1:1000; BJ11258993; Bioss, China), TNF-α (1:400; GB11188; Servicebio, China), IL-6 (1:1000; 21865–1-AP; Proteintech, China), MCP-1 (1:1500; ab25124; Abcam, USA), P65 (1:1000; 8242S; CST, Germany), Erk (1:1500; GB11560; Servicebio, China), P38 (1:1000;8690S; CST, Germany), JNK (1:1000; AF6318; Affinity, USA), p-P65 (1:1000; 3033S; CST, Germany), p-Erk (1:1000; AF1015; Affinity, USA), p-P38 (1:1000; 4511S; CST, Germany), p-JNK (1:1000;4668S; CST, Germany) and β-ACTIN (1:50000; 66009-l-Ig; Proteintech, China).

Techniques: Knockdown, Infection

Journal: Journal of Oral Microbiology

Article Title: MAT2A inhibition suppresses inflammation in Porphyromonas gingivalis -infected human gingival fibroblasts

doi: 10.1080/20002297.2023.2292375

Figure Lengend Snippet: NF-κB/MAPK pathway was involved in the enhanced MAT2A expression.

Article Snippet: Proteins at 10 μL per lane were separated by 4–20% SDS-PAGE (Beyotime, China) and then transferred onto PVDF membranes (Merck, Germany), which were blocked with 5% bovine albumin and then incubated with primary antibodies to MAT2A (1:1000; 55309–1-AP; Proteintech, China), IL-1β (1:1000; BJ11258993; Bioss, China), TNF-α (1:400; GB11188; Servicebio, China), IL-6 (1:1000; 21865–1-AP; Proteintech, China), MCP-1 (1:1500; ab25124; Abcam, USA), P65 (1:1000; 8242S; CST, Germany), Erk (1:1500; GB11560; Servicebio, China), P38 (1:1000;8690S; CST, Germany), JNK (1:1000; AF6318; Affinity, USA), p-P65 (1:1000; 3033S; CST, Germany), p-Erk (1:1000; AF1015; Affinity, USA), p-P38 (1:1000; 4511S; CST, Germany), p-JNK (1:1000;4668S; CST, Germany) and β-ACTIN (1:50000; 66009-l-Ig; Proteintech, China).

Techniques: Expressing