mpc 83 cells  (Dojindo Labs)

Journal: Cell & Bioscience

Article Title: Blockade of the Arid5a/IL-6/STAT3 axis underlies the anti-inflammatory effect of Rbpjl in acute pancreatitis

doi: 10.1186/s13578-022-00819-1

Figure Lengend Snippet: Rbpjl is under-expressed in both pancreatic tissues of AP mice and LPS-induced pancreatic acinar cells. A A volcano map of the expression of DEGs in AP samples in GSE121038 microarray (red dot indicates highly expressed genes, green dot indicates poorly expressed genes, X axis represents -log10, and Y axis indicates logFC. B Venn map of AP-related genes from GeneCards and CDT databases and differential genes obtained from GEO database. C A heat map of 32 candidate genes involved in the regulation of AP in AP samples (green to red indicates the expression value from small to large). D The expression of Rbpjl in AP samples in the GSE121038 microarray. E Immunohistochemistry analysis of Rbpjl protein in pancreatic tissues of sham-operated or AP mice (* p < 0.05 vs. sham-operated mice). F The apoptosis rate in pancreatic tissues of sham-operated or AP mice was detected by TUNEL staining (* p < 0.05 vs. sham-operated mice). G The expression of pro-inflammatory factors in the serum of sham-operated or AP mice was detected by ELISA (* p < 0.05 vs. sham-operated mice). H, Western blot assay was used to detect the protein expression of Rbpjl in control pancreatic acinar cells or pancreatic acinar cells treated with LPS at different time points (* p < 0.05 vs. control cells). I The expression of pro-inflammatory factors in control pancreatic acinar cells or pancreatic acinar cells treated with LPS for 24 h was detected by Western blot assay. (* p < 0.05 vs. control cells). The mouse experiments were performed 14 days after AP modeling, with n = 8 for mice in each group. The cell experiment was conducted three times independently

Article Snippet: Mouse pancreatic acinar cell line (MPC-83, CL-0518), procured from Procell (Wuhan, Hubei, China), was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (HyClone Company, Logan, UT), supplemented with 100 U/mL penicillin and 100 U/mL streptomycin (Gibco Company, Grand Island, NY) in a 5% CO 2 incubator at 37 °C.

Techniques: Expressing, Microarray, Immunohistochemistry, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: Cell & Bioscience

Article Title: Blockade of the Arid5a/IL-6/STAT3 axis underlies the anti-inflammatory effect of Rbpjl in acute pancreatitis

doi: 10.1186/s13578-022-00819-1

Figure Lengend Snippet: Overexpression of Rbpjl inhibits LPS-induced inflammatory response in pancreatic acinar cells. Control and LPS-induced MPC-83 cells were treated with oe-Rbpjl or sh-Rbpjl. A The protein expression of Rbpjl in control and LPS-induced MPC-83 cells was detected by Western blot assay. B The control and LPS-induced MPC-83 cell viability was detected by CCK-8 assay. C Apoptosis of control and LPS-induced MPC-83 cells detected by flow cytometry. D GSH and MDA production in control and LPS-induced MPC-83 cells were detected using an ultraviolet spectrophotometer. E The production of ROS in control and LPS-induced MPC-83 cells. F The expression of pro-inflammatory factors in control and LPS-induced MPC-83 cells was detected by Western blot assay. & p < 0.05 vs. control MPC-83 cells + oe-NC. @ p < 0.05 vs. control MPC-83 cells + sh-NC. * p < 0.05 vs. MPC-83 cells treated with LPS + oe-NC. # p < 0.05 vs. MPC-83 cells treated with LPS + sh-NC. Cell experiments were performed after 72 h of lentivirus transduction and 24 h of LPS stimulation. The cell experiment was conducted three times independently

Article Snippet: Mouse pancreatic acinar cell line (MPC-83, CL-0518), procured from Procell (Wuhan, Hubei, China), was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (HyClone Company, Logan, UT), supplemented with 100 U/mL penicillin and 100 U/mL streptomycin (Gibco Company, Grand Island, NY) in a 5% CO 2 incubator at 37 °C.

Techniques: Over Expression, Control, Expressing, Western Blot, CCK-8 Assay, Flow Cytometry, Spectrophotometry, Transduction

Journal: Cell & Bioscience

Article Title: Blockade of the Arid5a/IL-6/STAT3 axis underlies the anti-inflammatory effect of Rbpjl in acute pancreatitis

doi: 10.1186/s13578-022-00819-1

Figure Lengend Snippet: Rbpjl down-regulates the expression of Arid5a by binding to its promoter region. A Analysis of the expression of Arid5a in AP samples in the GSE121038 dataset. B The binding site of Rbpjl and the Arid5a promoter region was predicted by the JASPAR website. C Detection of mRNA and protein expression of Arid5a in the pancreatic tissues of AP mice by RT-qPCR and Western blot assay (* p < 0.05 vs. sham-operated mice). D Detection of mRNA and protein expression of Arid5a in the LPS-induced MPC-83 cells at different time points by RT-qPCR and Western blot assay (* p < 0.05 vs. control cells). E Correlation of Arid5a protein expression with Rbpjl protein expression in LPS-induced MPC-83 cells at different time points analyzed by Pearson's correlation coefficient (* p < 0.0001). F The binding of Rbpjl to the Arid5a promoter region was analyzed by ChIP after pancreatic acinar cells with oe-Rbpjl or sh-Rbpjl (for 72 h) were stimulated by LPS (for 24 h) (* p < 0.05 vs. cells treated with sh-NC. # p < 0.05 vs. cells treated with oe-NC). G, Rbpjl protein binding to the Arid5a promoter region in pancreatic acinar cells was analyzed by EMSA. H Luciferase activity of WT-Arid5a promoter and MUT-Arid5a promoter in HEK-293 T cells transfected with oe-Rbpjl was detected by dual luciferase reporter assay (* p < 0.05 vs. HEK-293 T cells transfected with oe-NC). I The mRNA and protein expression of Arid5a was measured by RT-qPCR and Western blot assay after pancreatic acinar cells with oe-Rbpjl or sh-Rbpjl (for 72 h) were stimulated by LPS (for 24 h) (* p < 0.05 vs. cells treated with sh-NC. # p < 0.05 vs. cells treated with oe-NC). The mouse experiments were performed 14 days after AP modeling, with n = 8 for mice in each group. The cell experiment was conducted three times independently

Article Snippet: Mouse pancreatic acinar cell line (MPC-83, CL-0518), procured from Procell (Wuhan, Hubei, China), was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (HyClone Company, Logan, UT), supplemented with 100 U/mL penicillin and 100 U/mL streptomycin (Gibco Company, Grand Island, NY) in a 5% CO 2 incubator at 37 °C.

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Western Blot, Control, Protein Binding, Luciferase, Activity Assay, Transfection, Reporter Assay

Journal: Cell & Bioscience

Article Title: Blockade of the Arid5a/IL-6/STAT3 axis underlies the anti-inflammatory effect of Rbpjl in acute pancreatitis

doi: 10.1186/s13578-022-00819-1

Figure Lengend Snippet: Arid5a activates the IL-6/STAT3 axis in pancreatic acinar cells. A The interaction network of Arid5a in the STRING website. B The interaction network of IL-6 in the STRING website. C Detection of intranuclear protein expression of STAT3 and STAT3 phosphorylation level in the pancreatic tissues of AP mice and LPS-induced MPC-83 cells (for 6 h) by Western blot assay (* p < 0.05 vs. sham-operated mice. # p < 0.05 vs. control MPC-83 cells). D The expression of intranuclear STAT3 protein, Arid5a, total protein expression of STAT3 as well as STAT3 phosphorylation level in the LPS-induced MPC-83 cells (for 6 h) treated with sh-Arid5a was measured by Western blot assay (* p < 0.05 vs. LPS-induced MPC-83 cells treated with sh-NC). E The expression of pro-inflammatory factor IL-6 in LPS-induced MPC-83 cells (for 6 h) treated with sh-Arid5a was detected by RT-qPCR (* p < 0.05 vs. LPS-induced MPC-83 cells treated with sh-NC). The mouse experiments were performed 14 days after AP modeling, with n = 8 for mice in each group. The cell experiment was conducted three times independently

Article Snippet: Mouse pancreatic acinar cell line (MPC-83, CL-0518), procured from Procell (Wuhan, Hubei, China), was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (HyClone Company, Logan, UT), supplemented with 100 U/mL penicillin and 100 U/mL streptomycin (Gibco Company, Grand Island, NY) in a 5% CO 2 incubator at 37 °C.

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Quantitative RT-PCR

Journal: Cell & Bioscience

Article Title: Blockade of the Arid5a/IL-6/STAT3 axis underlies the anti-inflammatory effect of Rbpjl in acute pancreatitis

doi: 10.1186/s13578-022-00819-1

Figure Lengend Snippet: Rbpjl downregulates Arid5a expression and thus inhibits the IL-6/STAT3 axis to alleviate inflammatory response in LPS-induced pancreatic acinar cells. A mRNA expression of Rbpjl, Arid5a and IL-6 after MPC-83 cells with oe-Rbpjl or combined with oe-Arid5a (for 72 h) were stimulated by LPS (for 24 h) determined by RT-qPCR. B Western blot assay was used to detect the expression of Rbpjl, Arid5a and intranuclear STAT3 proteins, total protein expression of STAT3 as well as STAT3 phosphorylation level after MPC-83 cells with oe-Rbpjl or combined with oe-Arid5a (for 72 h) were stimulated by LPS (for 24 h). C CCK-8 assay was used to detect the proliferation of MPC-83 cells with oe-Rbpjl or combined with oe-Arid5a (for 72 h) stimulated by LPS (for 24 h). D CCK-8 assay was used to detect the proliferation of MPC-83 cells with oe-Arid5a (for 72 h) or combined with JSI-124 (for 24 h) stimulated by LPS (for 24 h). E Apoptosis of MPC-83 cells with oe-Rbpjl or combined with oe-Arid5a (for 72 h) stimulated by LPS (for 24 h) detected by flow cytometry. F Apoptosis of MPC-83 cells with oe-Arid5a (for 72 h) or combined with JSI-124 (for 24 h) stimulated by LPS (for 24 h) detected by flow cytometry. G Ultraviolet spectrophotometers were used to detect GSH and MDA production in MPC-83 cells with oe-Rbpjl or combined with oe-Arid5a (for 72 h) stimulated by LPS (for 24 h). H Ultraviolet spectrophotometers were used to detect GSH and MDA production in MPC-83 cells with oe-Arid5a (for 72 h) or combined with JSI-124 (for 24 h) stimulated by LPS (for 24 h). I TNF-α, IL-1β and IL-6 expression in MPC-83 cells with oe-Rbpjl or combined with oe-Arid5a (for 72 h) stimulated by LPS (for 24 h). J TNF-α, IL-1β and IL-6 expression in MPC-83 cells with oe-Arid5a (for 72 h) or combined with JSI-124 (for 24 h) stimulated by LPS (for 24 h). * p < 0.05 vs. MPC-83 cells treated with LPS + oe-NC. # p < 0.05 vs. MPC-83 cells treated with LPS + oe-Rbpjl + oe-NC. & p < 0.05 vs. MPC-83 cells treated with LPS + oe-Arid5a + DMSO. The cell experiment was conducted three times independently

Article Snippet: Mouse pancreatic acinar cell line (MPC-83, CL-0518), procured from Procell (Wuhan, Hubei, China), was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (HyClone Company, Logan, UT), supplemented with 100 U/mL penicillin and 100 U/mL streptomycin (Gibco Company, Grand Island, NY) in a 5% CO 2 incubator at 37 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Phospho-proteomics, CCK-8 Assay, Flow Cytometry

Journal: Cell & Bioscience

Article Title: Blockade of the Arid5a/IL-6/STAT3 axis underlies the anti-inflammatory effect of Rbpjl in acute pancreatitis

doi: 10.1186/s13578-022-00819-1

Figure Lengend Snippet: Rbpjl alleviates AP by downregulating Arid5a and blocking the IL-6/STAT3 axis in vivo. A Rbpjl and Arid5a protein expression in pancreatic tissues of mice treated with oe-Rbpjl or combined with oe-Arid5a determined by Western blot assay. B The intranuclear STAT3 protein expression, total protein expression of STAT3 as well as STAT3 phosphorylation level in pancreatic tissues of mice treated with oe-Rbpjl or combined with oe-Arid5a were measured by Western blot assay. C HE staining was used to observe the degree of pancreatic injury, and the degree of pancreatic injury was graded and evaluated in mice treated with oe-Rbpjl or combined with oe-Arid5a. D The weight ratio of pancreatic tissues to total body in mice treated with oe-Rbpjl or combined with oe-Arid5a was detected. E TUNEL staining was used to detect the apoptosis rate in pancreatic tissues of mice treated with oe-Rbpjl or combined with oe-Arid5a. F The expression of pro-inflammatory factors in the serum of mice treated with oe-Rbpjl or combined with oe-Arid5a was detected by ELISA. The mouse experiments were performed 14 days after AP modeling, with n = 8 for mice in each group

Article Snippet: Mouse pancreatic acinar cell line (MPC-83, CL-0518), procured from Procell (Wuhan, Hubei, China), was cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) (HyClone Company, Logan, UT), supplemented with 100 U/mL penicillin and 100 U/mL streptomycin (Gibco Company, Grand Island, NY) in a 5% CO 2 incubator at 37 °C.

Techniques: Blocking Assay, In Vivo, Expressing, Western Blot, Phospho-proteomics, Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay