Journal: Pharmaceutical Biology

Article Title: Salvianolic acid B attenuates tubulointerstitial fibrosis by inhibiting EZH2 to regulate the PTEN/Akt pathway

doi: 10.1080/13880209.2022.2148169

Figure Lengend Snippet: SAB inhibits renal fibrosis through inhibiting EZH2 and reversing the low level of PTEN and phosphorylation of AKT increase in obstructed kidneys. (A) Representative images of PTEN expression detected by IHC (Magnification × 400, Bar = 50 μm). (B) Quantitative analysis of PTEN positive area ( n = 6). (C) Immunoblot analysis of p-AKT and AKT. (D) Immunoblot analysis of EZH2 and H3k27me3. (E) The ratio of AKT to GAPDH protein and phosphorylated AKT to AKT protein was measured ( n = 6). (F) The ratio of EZH2, H3k27me3 to GAPDH protein was measured ( n = 6). ## p < 0.01 vs. vehicle-sham vs. vehicle-sham; * p < 0.05 vs. vehicle-UUO; ** p < 0.01 vs. vehicle-UUO.

Article Snippet: For immunohistochemical staining, the 4 μm sections were incubated with 1:200 diluted PTEN rabbit Anti-body (9559S, CST, USA).

Techniques: Expressing, Western Blot

Journal: Pharmaceutical Biology

Article Title: Salvianolic acid B attenuates tubulointerstitial fibrosis by inhibiting EZH2 to regulate the PTEN/Akt pathway

doi: 10.1080/13880209.2022.2148169

Figure Lengend Snippet: SAB alleviates renal fibrosis through inhibiting EZH2 and reversing the low level of PTEN and phosphorylation of AKT increase in AAN model kidneys. (A) Representative images of PTEN expression detected by IHC (magnification × 400, Bar = 50 μm). (B) Quantitative analysis of PTEN positive area ( n = 6). (C) Immunoblot analysis of p-AKT and AKT. (D) Immunoblot analysis of EZH2 and H3k27me3. (E) The ratio of AKT to GAPDH protein and phosphorylated AKT to AKT protein was measured ( n = 6). (F) The ratio of EZH2, H3k27me3 to GAPDH protein was measured ( n = 6). ## p < 0.01 vs. vehicle-control; * p < 0.05 vs. vehicle-AAN; ** p < 0.01 vs. vehicle-AAN.

Article Snippet: For immunohistochemical staining, the 4 μm sections were incubated with 1:200 diluted PTEN rabbit Anti-body (9559S, CST, USA).

Techniques: Expressing, Western Blot

Journal: Journal of Inflammation (London, England)

Article Title: Rho iso-alpha acids from hops inhibit the GSK-3/NF-κB pathway and reduce inflammatory markers associated with bone and cartilage degradation

doi: 10.1186/1476-9255-6-26

Figure Lengend Snippet: Effect of RIAA on the inhibition of GSK3 pathway . (A) GSK-3α was incubated with RIAA (0, 5, 25 and 50 μg/ml) and the kinase activity was determined. Data represent 1 representative experiment and are expressed as % activity. (B) GSK-3β was incubated with RIAA (0, 1, 5, 25 and 50 μg/ml) and the kinase activity was determined. Data represent 1 representative experiment and are expressed as % activity. (C) RAW 264.7 cells were incubated with RIAA (0, 1, 5, 10 and 20 μg/ml), or GSK-3 inhibitors LiCl (10 mM) and SB216763 (10 μM) for 1 h. Cell lysates were analyzed for the inhibition of phosphorylation of β-catenin by western blot.

Article Snippet: Phospho-ERK1/2, phospho-p38, phospho-JNK, phospho-β-catenin anti-bodies were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Inhibition, Incubation, Activity Assay, Western Blot

Journal: Journal of Biomedical Science

Article Title: Cytoplasmic p21 induced by p65 prevents doxorubicin-induced cell death in pancreatic carcinoma cell line

doi: 10.1186/1423-0127-19-15

Figure Lengend Snippet: Cytoplasmic p21 induced by p65 is necessary to prevent the activation of procaspase-3 in PANC1 cell . Cells were transiently transfected with or without over-expression of p65 and p65-targeted shRNA followed by DOX treatment (2 μg/ml) for 24 h. (A) The level of p65 protein was detected by Western blotting. (B) Cell survival was examined by CCK8 kit. Error bar indicates the standard error of the mean of three independent experiments. (C)The nuclear or cytoplasmic p21 protein (above) and pro-caspase-3 or active caspase-3 (bottom) were detected by Western blotting. GAPDH was used as controls.

Article Snippet: For immunoprecipitation, a 500 μg aliquot of cell lysate was incubated for 1 h at 4°C with the anti-p21 anti-body (2946, Cell Signaling).

Techniques: Activation Assay, Transfection, Over Expression, shRNA, Western Blot

Journal: Journal of Biomedical Science

Article Title: Cytoplasmic p21 induced by p65 prevents doxorubicin-induced cell death in pancreatic carcinoma cell line

doi: 10.1186/1423-0127-19-15

Figure Lengend Snippet: Pro-caspase-3 physically associated with cytoplasmic p21 induction by p65 in PANC1 cells . (A) Cells were transiently transfected with or without over-expression of p65 and p65-targeted shRNA followed by DOX treatment (2 μg/ml) for 24 h. Whole cell extract was subjected to immunoprecipitation with anti-p21 antibody and then immunoblotted with anti-pro-caspase-3 or -active-caspase-3 antibody. (B) Cells were transfected with vector expressing wild-type (1-585), N-terminal (1-372) and C-terminal (373-585) region of p21 followed by DOX treatment (2 μg/ml) for 24 h. Immunoprecipitation with an antibody against myc peptide. The pro-caspase-3 and p21 amounts were assessed by Western blotting.

Article Snippet: For immunoprecipitation, a 500 μg aliquot of cell lysate was incubated for 1 h at 4°C with the anti-p21 anti-body (2946, Cell Signaling).

Techniques: Transfection, Over Expression, shRNA, Immunoprecipitation, Plasmid Preparation, Expressing, Western Blot

Journal: Journal of Biomedical Science

Article Title: Cytoplasmic p21 induced by p65 prevents doxorubicin-induced cell death in pancreatic carcinoma cell line

doi: 10.1186/1423-0127-19-15

Figure Lengend Snippet: Induction of p65 enhanced the p53-mediated cell death response to DOX in PANC1 cells . Cells were transiently transfected with or without over-expression of p53 alone or p53 and p65 together followed by DOX treatment (2 μg/ml) for 24 h. (A) The level of p53 or p65 protein was detected by Western blotting. (B) Cell survival was examined by CCK8 kit. Error bar indicates the standard error of the mean of three independent experiments. (C)The nuclear or cytoplasmic p21 protein (above) or pro-caspase-3 (bottom) were detected by Western blotting. GAPDH was used as controls. Whole cell extract was subjected to immunoprecipitation with anti-p21 antibody and then immunoblotted with anti-pro-caspase-3 antibody (middle). (D) Active-caspase-3 and active-caspase-8 protein were detected by Western blotting. GAPDH was used as controls.

Article Snippet: For immunoprecipitation, a 500 μg aliquot of cell lysate was incubated for 1 h at 4°C with the anti-p21 anti-body (2946, Cell Signaling).

Techniques: Transfection, Over Expression, Western Blot, Immunoprecipitation