Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Interaction analysis of the control drug Gemcitabine and the proposed nutraceutical therapeutics Queuine and Thiamine with the MAP4K4 binding site. ( A ) 3D binding pose of Gemcitabine (left) and 2D representation (right) of Gemcitabine docked in the MAP4K4 binding site. ( B ) 3D and 2D binding pose of Queuine ( C ) 3D and 2D binding pose of Thiamine. Coloring schemes: hydrogen bonding (purple arrows), pi-pi stacking (green line), and pi-cation interactions (red line)

Article Snippet: The ELISA was performed following the protocol provided by the Human Mitogen-activated Protein Kinase 4 (MAP4K4) ELISA Kit, which is intended for the quantitative detection of MAP4K4 (BT Lab, Cat. no. E5689Hu) with slight modifications.

Techniques: Control, Binding Assay

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Protein-ligand complex root mean square deviation (RMSD) ( A ) and root mean square fluctuation (RMSF) ( B ) plots for the apo, Gemcitabine, Queuine, and Thiamine bound forms of the MAP4K4 protein

Article Snippet: The ELISA was performed following the protocol provided by the Human Mitogen-activated Protein Kinase 4 (MAP4K4) ELISA Kit, which is intended for the quantitative detection of MAP4K4 (BT Lab, Cat. no. E5689Hu) with slight modifications.

Techniques:

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Effects of different concentrations of Queuine and Gemcitabine on MAP4K4 activity. Absorbance values on the y-axis represent the enzyme amount measured using an ELISA kit, with different concentrations of Queuine ( A ) and Gemsitabine ( B ) indicated on the x-axis. The differences between the bar pairs marked with * ( p < 0.05) and *** ( p < 0.001) are significant

Article Snippet: The ELISA was performed following the protocol provided by the Human Mitogen-activated Protein Kinase 4 (MAP4K4) ELISA Kit, which is intended for the quantitative detection of MAP4K4 (BT Lab, Cat. no. E5689Hu) with slight modifications.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC Biotechnology

Article Title: In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer

doi: 10.1186/s12896-025-01033-w

Figure Lengend Snippet: Synergetic effects of Queuine and Gemcitabine on MAP4K4 activity. Absorbance values represent the enzyme amount measured using an ELISA kit. The difference between bar pairs marked with *** are significant at p < 0.001

Article Snippet: The ELISA was performed following the protocol provided by the Human Mitogen-activated Protein Kinase 4 (MAP4K4) ELISA Kit, which is intended for the quantitative detection of MAP4K4 (BT Lab, Cat. no. E5689Hu) with slight modifications.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Signal transduction and targeted therapy

Article Title: IGF-1R inhibition induces MEK phosphorylation to promote survival in colon carcinomas.

doi: 10.1038/s41392-020-0204-0

Figure Lengend Snippet: Fig. 5 Inactivation of MEK1 or MEK2 stimulates AKT phosphorylation. a, b Knockdown of MEK1 or MEK2 boosts AKT phosphorylation. Cells were transfected with scrambled siRNA, mek1 siRNA, or mek2 siRNA. After 48 h, cell lysates were subjected to western blot analysis. a Representative images are shown. b Densitometric quantification of phospho-AKT levels in (a). The ratio of phospho-AKT/GAPDH of transfection with scrambled siRNA is designated as 1. Data from three independent experiments were analyzed by one-sample t-test (mean ± SD; *P < 0.05). c, d Inhibition of MEK1/2 activation induces AKT phosphorylation. HCT116 cells were treated with 0, 1, 5, and 10 μM of U0126 for 72 h. The cell lysates were used to examine the levels of phospho-AKT. c Representative western blot images are shown. b Densitometric quantification of phospho-AKT and phospho-p70S6K1 levels in (c). The ratio of phospho-AKT/GAPDH or phospho-p70S6K1 in cells with 0 μM of U0126 is designated as 1. Data from three independent experiments were analyzed by one-sample t-test (mean ± SD; *P < 0.05; **P < 0.01)

Article Snippet: The MEK2 (67410–1-Ig) antibody was purchased from Proteintech (1:4,000 dilution).

Techniques: Phospho-proteomics, Knockdown, Transfection, Western Blot, Inhibition, Activation Assay

Journal: Journal of Inflammation Research

Article Title: Curcumin Alleviates Osteoarthritis Through the p38MAPK Pathway: Network Pharmacological Prediction and Experimental Confirmation

doi: 10.2147/jir.s459867

Figure Lengend Snippet: Figure 5 Western blot analysis of curcumin regulation of the p38MAPK pathway. (A–C) Western blot and quantitative analysis of p38 and p-p38. (D–F) Western blot and quantitative analysis of ASK1 and MEKK3. (G and H) Western blot images and quantitative analysis of PKCδ and p-PKCδ. (I–L) Western blot and quantitative analysis of BCL-2 and caspase-3. (n = 3). *p < 0.05,** p < 0.01,***p < 0.001.

Article Snippet: Antibodies against Caspase-3, Bcl-2, PKCδ, MEKK3, ASK1, and GAPDH were purchased from Boster (Wuhan, China).

Techniques: Western Blot

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 expression of MAP2K3 in clinic human HCC and matched adjacent non-tumor tissues was evaluated by immunohistochemistry staining using rabbit anti-MAP2K3 antibody (1:100, Boster, Wuhan, China).

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 expression of MAP2K3 in clinic human HCC and matched adjacent non-tumor tissues was evaluated by immunohistochemistry staining using rabbit anti-MAP2K3 antibody (1:100, Boster, Wuhan, China).

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 expression of MAP2K3 in clinic human HCC and matched adjacent non-tumor tissues was evaluated by immunohistochemistry staining using rabbit anti-MAP2K3 antibody (1:100, Boster, Wuhan, China).

Techniques: Infection, Plasmid Preparation, Expressing, Western Blot