Journal: BMC Cancer

Article Title: MicroRNA-21 promotes hepatocellular carcinoma HepG2 cell proliferation through repression of mitogen-activated protein kinase-kinase 3

doi: 10.1186/1471-2407-13-469

Figure Lengend Snippet: The expression of MAP2K3 in human HCC tissues determined by IHC

Article Snippet: The membranes were probed with rabbit anti-MAP2K3 antibody and anti-GAPDH antibody (Boster, Wuhan, China) or (1:200, Boster, Wuhan, China) were for the interested protein MAP2K3 and endogenous GAPDH for loading control, respectively.

Techniques: Expressing, Immunohistochemistry

Journal: BMC Cancer

Article Title: MicroRNA-21 promotes hepatocellular carcinoma HepG2 cell proliferation through repression of mitogen-activated protein kinase-kinase 3

doi: 10.1186/1471-2407-13-469

Figure Lengend Snippet: Validation of MAP2K3 mRNA as a target of miR-21. (A) : Sequence of potential binding site of miR-21 in the 3’UTR of MAP2K3 mRNA (top panel), mutations were introduced into the binding site for generation of mutated MAP2K3 3’TUR (bottom panel). ( B and C ): Validation of miR-21 target using MAP2K3 3’UTR luciferase reporter. Cells co-transfected with pMIR-Report/MAP2K3 3’UTR (WT) or pMIR-Report/Mut-MAP2K3 3’UTR (Mut) and pAd/pri-miR-21 (B) , pAd/miR-21/inhibitor (C) , and pAd/con plasmids showed a decreased luciferase activity in pAd/pri-miR-21 cells (B) . Luciferase activity after site directed mutagenesis of the 3’UTR of MAP2K3 mRNA in the miR-21 seed sequence (pMIR-Report/Mut-MAP2K3) was significantly higher with respect to the pMIR-Report/MAP2K3 vector transfected cells ( B and C ). Results represented the mean ± SD from three independent triplicated experiments (N=9).

Article Snippet: The membranes were probed with rabbit anti-MAP2K3 antibody and anti-GAPDH antibody (Boster, Wuhan, China) or (1:200, Boster, Wuhan, China) were for the interested protein MAP2K3 and endogenous GAPDH for loading control, respectively.

Techniques: Biomarker Discovery, Sequencing, Binding Assay, Luciferase, Transfection, Activity Assay, Mutagenesis, Plasmid Preparation

Journal: BMC Cancer

Article Title: MicroRNA-21 promotes hepatocellular carcinoma HepG2 cell proliferation through repression of mitogen-activated protein kinase-kinase 3

doi: 10.1186/1471-2407-13-469

Figure Lengend Snippet: miR-21 targets MAP2K3 mRNA. The HepG2 cells were infected with Ad/pri-miR-21, Ad/miR-21/inhibitor or Ad/con adenoviral vector. The expression of MAP2K3 was detected by immunoblotting analysis against anti-MAP2K3 antibody. Compared with Ad/con group, *: p <0.05. Data in A represented the mean ± SD from three independent triplicated experiments (N=9).

Article Snippet: The membranes were probed with rabbit anti-MAP2K3 antibody and anti-GAPDH antibody (Boster, Wuhan, China) or (1:200, Boster, Wuhan, China) were for the interested protein MAP2K3 and endogenous GAPDH for loading control, respectively.

Techniques: Infection, Plasmid Preparation, Expressing, Western Blot

Journal: Journal of Dental Sciences

Article Title: Overexpression of sprouty 1 protein in human oral squamous cell carcinogenesis

doi: 10.1016/j.jds.2020.07.013

Figure Lengend Snippet: Western blot analyses: phosphor-ERK and total-ERK in human oral cancer cell lines as compared with a primary culture of normal oral mucosa (HOK). Upregulation of phosphor-ERK (normalized to total-ERK) in OECM1 and Ca922 cell lines as compared with HOK, and a slightly decreased expression in SAS as compared with HOK. Results were quantified using densitometric analysis, normalized to the level of β-actin, and expressed as a fold change relative to the normal oral mucosa. Bars represent means ± standard deviation of the mean (∗ P < 0.05). A representative result of three independent experiments is shown.

Article Snippet: Further, samples were analyzed using 10% SDS-PAGE (Sigma-Aldrich) gels, and the proteins were transmitted onto a PVDF membrane (Sigma-Aldrich) using Bio-Rad's transblot with primary antibodies against phosphor-ERK (Boster Biological Technology, CA, USA; Cat. No. P00104; 1:1000) and total-ERK (Boster Biological Technology; Cat. No. P00104; 1:1000), with species specificity for human tissues and an observed molecular weight of 42–44 kDa; and β-actin (Sigma-Aldrich; 1:1000), followed by horseradish peroxidase (HRP)-conjugated secondary antibodies (Sigma-Aldrich; 1:5000).

Techniques: Western Blot, Expressing, Standard Deviation

Journal:

Article Title: Hyperosmotic Stress Induces Nuclear Factor-?B Activation and Interleukin-8 Production in Human Intestinal Epithelial Cells

doi:

Figure Lengend Snippet: Hyperosmolarity induces p38 and p42/44 activation in HT-29 cells (A). Cells were incubated with hyperosmotic medium prepared by the addition of mannitol (for a final concentration of 100 mmol/L) to isosmolar medium for the indicated time periods. p38 and p42/44 activation was determined using Western blotting using antibodies raised against the active, double-phosphorylated form of p38 and p42/44. This figure is representative of two separate experiments. Treatment of HT-29 cells with the selective p38 inhibitor SB203580 or selective p42/44 inhibitor PD98059 suppresses the hyperosmolarity-induced IL-8 response (B). Hyperosmolarity was achieved by the addition of mannitol (for a final concentration of 100 mmol/L) to isosmolar medium. In control wells, cells were incubated with isosmolar medium. Data are mean ± SEM of n = 12 wells from two separate experiments. *, P < 0.05; **, P < 0.01. Dotted bar, no mannitol; cross-hatched bars, mannitol.

Article Snippet: The membrane was probed with anti-phospho-p38 or anti-phospho-p42/44 MAP kinase antibody (Promega) and subsequently incubated with a secondary horseradish peroxidase-conjugated donkey anti-rabbit antibody (Boehringer).

Techniques: Activation Assay, Incubation, Concentration Assay, Western Blot