Journal: Molecular Cell

Article Title: The ribotoxic stress response drives acute inflammation, cell death, and epidermal thickening in UV-irradiated skin in vivo

doi: 10.1016/j.molcel.2024.10.044

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ZAK , Proteintech , Cat#14945-1-AP; RRID: AB_1064269.

Techniques: Recombinant, Reverse Transcription, In Situ, Knock-Out, Variant Assay, Software, Purification, Blocking Assay

Journal: Cells

Article Title: Intracellular Angiotensin II Stimulation of Sodium Transporter Expression in Proximal Tubule Cells via AT 1 (AT 1a ) Receptor-Mediated, MAP Kinases ERK1/2- and NF-кB-Dependent Signaling Pathways

doi: 10.3390/cells12111492

Figure Lengend Snippet: The roles of the MAP kinase and NF-κB signaling pathways in mediating ECFP/Ang II-induced NHE3 expression in wild-type and Agtr1a -/- mPCT cells. Panel ( A ) shows that in wild-type mPCT cells, ECFP/Ang II stimulated NHE3 expression significantly, and the response was attenuated by the MEK1/MEK2 kinase inhibitor U0126 and the NF-κB activation inhibitor Ro 106–9920, respectively. However, the MEK inhibitor PD 980659 and the p38 MAP kinase inhibitor SB202196 failed to attenuate the effect of ECFP/Ang II on NHE3 expression. Panel ( B ) shows that in Agtr1a -/- mPCT cells, ECFP/Ang II failed to stimulate NHE3 expression, and the inhibitors of the MAP kinases and NF-κB signaling pathways had no significant effects on NHE3 expression. ** p < 0.01 vs. control WT mPCT cells. ++ p < 0.01 vs. WT mPCT cells transfected with ECFP/Ang II.

Article Snippet: To determine the potential signaling mechanisms involved in Ad- Sglt2-ECFP/Ang II -induced biological responses, WT and Agtr1a -/- mPCT cells expressing Ad- Sglt2-ECFP/Ang II were concurrently treated with the AT 1 receptor antagonist losartan (10 μM; Tocris, Minneapolis, MN, USA), the AT 2 receptor antagonist PD 123319 (10 μM; Tocris, Minneapolis, MN, USA), the MEK1/MEK2 kinase inhibitor U0126 (1 μM; Tocris, Minneapolis, MN, USA), the MEK inhibitor PD 980659 (1 μM; Tocris, Minneapolis, MN, USA), the NF-κB activation inhibitor RO 106–9920 (10 μM; Tocris, Minneapolis, MN, USA), and the p38 MAP kinase inhibitor SB202196 (10 μM; MCE, Belleville, NJ, USA).

Techniques: Protein-Protein interactions, Expressing, Activation Assay, Control, Transfection

Journal: Cells

Article Title: Intracellular Angiotensin II Stimulation of Sodium Transporter Expression in Proximal Tubule Cells via AT 1 (AT 1a ) Receptor-Mediated, MAP Kinases ERK1/2- and NF-кB-Dependent Signaling Pathways

doi: 10.3390/cells12111492

Figure Lengend Snippet: The roles of AT 1 and AT 2 receptors, the MAP kinases, and NF-κB signaling pathways in mediating ECFP/Ang II-induced Na + /HCO 3 - cotransporter expression in wild-type mPCT cells. Panel ( A ) shows that ECFP/Ang II significantly increased Na + /HCO 3 - expression, and the response was attenuated by losartan but not by PD123319, suggesting a dominant role of AT 1 receptors in mPCT cells. Panel ( B ) shows that the MEK1/MEK2 kinase inhibitor U0126, the NF-κB activation inhibitor Ro 106–9920, and the MEK inhibitor PD 980659 attenuated the effects of ECFP/Ang II on expression, but the p38 MAP kinase inhibitor SB202196 had no effect on Na + /HCO 3 - expression. ** p < 0.01 vs. control WT mPCT cells. ++ p < 0.01 vs. WT mPCT cells transfected with ECFP/Ang II.

Article Snippet: To determine the potential signaling mechanisms involved in Ad- Sglt2-ECFP/Ang II -induced biological responses, WT and Agtr1a -/- mPCT cells expressing Ad- Sglt2-ECFP/Ang II were concurrently treated with the AT 1 receptor antagonist losartan (10 μM; Tocris, Minneapolis, MN, USA), the AT 2 receptor antagonist PD 123319 (10 μM; Tocris, Minneapolis, MN, USA), the MEK1/MEK2 kinase inhibitor U0126 (1 μM; Tocris, Minneapolis, MN, USA), the MEK inhibitor PD 980659 (1 μM; Tocris, Minneapolis, MN, USA), the NF-κB activation inhibitor RO 106–9920 (10 μM; Tocris, Minneapolis, MN, USA), and the p38 MAP kinase inhibitor SB202196 (10 μM; MCE, Belleville, NJ, USA).

Techniques: Protein-Protein interactions, Expressing, Activation Assay, Control, Transfection

Journal: Cells

Article Title: Intracellular Angiotensin II Stimulation of Sodium Transporter Expression in Proximal Tubule Cells via AT 1 (AT 1a ) Receptor-Mediated, MAP Kinases ERK1/2- and NF-кB-Dependent Signaling Pathways

doi: 10.3390/cells12111492

Figure Lengend Snippet: The roles of AT 1 and AT 2 receptors, the MAP kinases, and NF-κB signaling pathways in mediating ECFP/Ang II-induced NF-κB, p65 expression in wild-type and Agtr1a -/- mPCT cells. Panel ( A ) shows that ECFP/Ang II increased NF-κB, p65 expression in wild-type mPCT cells, and the response was attenuated by both losartan and PD123319, supporting an important role of AT 1 and AT 2 receptors in mediating ECFP/Ang II-induced NF-κB, p65 expression in wild-type mPCT cells. Panel ( B ) shows that ECFP/Ang II alone had no significant effect on NF-κB, p65 expression in Agtr1a -/- mPCT cells, but both losartan and PD123319 potentiated this response. Panel ( C ) shows that in wild-type mPCT cells, the effect of ECFP/Ang II on NF-κB, p65 expression was attenuated by the MEK1/MEK2 kinase inhibitor U0126, the NF-κB activation inhibitor Ro 106–9920, and the MEK inhibitor PD 980659, respectively. However, the p38 MAP kinase inhibitor SB202196 had no effect on ECFP/Ang II-induced NF-κB, p65 expression in wild-type mPCT cells. ** p < 0.01 vs. control WT or Agtr1a -/- mPCT cells. ++ p < 0.01 vs. WT mPCT cells transfected with ECFP/Ang II, or Agtr1a -/- mPCT cells transfected with ECFP/ANG II.

Article Snippet: To determine the potential signaling mechanisms involved in Ad- Sglt2-ECFP/Ang II -induced biological responses, WT and Agtr1a -/- mPCT cells expressing Ad- Sglt2-ECFP/Ang II were concurrently treated with the AT 1 receptor antagonist losartan (10 μM; Tocris, Minneapolis, MN, USA), the AT 2 receptor antagonist PD 123319 (10 μM; Tocris, Minneapolis, MN, USA), the MEK1/MEK2 kinase inhibitor U0126 (1 μM; Tocris, Minneapolis, MN, USA), the MEK inhibitor PD 980659 (1 μM; Tocris, Minneapolis, MN, USA), the NF-κB activation inhibitor RO 106–9920 (10 μM; Tocris, Minneapolis, MN, USA), and the p38 MAP kinase inhibitor SB202196 (10 μM; MCE, Belleville, NJ, USA).

Techniques: Protein-Protein interactions, Expressing, Activation Assay, Control, Transfection

Journal: Biochemistry and Biophysics Reports

Article Title: Bifendate inhibits cell PARthanatos by activating the MEK/ERK pathway

doi: 10.1016/j.bbrep.2026.102454

Figure Lengend Snippet: DDB sustains cellular phospho-ERK levels. (A–B) HeLa cells were pre-treated with DDB for 24 h, exposed to MNNG (60 μM, 15 min) and cultured for a further 8 h. ERK1 and phospho-ERK1/2 expression were analysed by immunoblot (A) and densitometry (B). (C) Cells pre-treated with DDB for 24 h were challenged with MNNG (60 μM, 15 min) and harvested 4 h later; MEK1/2 and phospho-MEK1 levels were analysed by immunoblot. (D) The thermal stability of MEK and ERK after 24 h pretreatment with DDB (25 μM) or DMSO was detected by Western blot. (E) The grey values of each protein band were analysed quantitatively. (n = 3). The values are expressed as the means ± SDs. Statistical analysis was performed using T-test. *p < 0.05. (F) The thermal stability of DUSP1 after 24 h pretreatment with DDB (25 μM) or DMSO was detected by Western blot. (G) The grey values of protein band were analysed quantitatively. (n = 3). The values are expressed as the means ± SDs. Statistical analysis was performed using T-test. (H) Cells were pre-treated or not with DDB for 24 h, exposed to MNNG (60 μM, 15 min) and cultured for 4 h then phospho-Bad expression was analysed. (I) Mitochondrial membrane potential was assessed by JC-1 staining after the indicated treatments. Scale bar: 50 μm. The values are expressed as the means ± SDs (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey's test. **p < 0.01.

Article Snippet: After sealing with 5 % skimmed milk, the membrane was incubated successively with the primary antibody (PAR/pADPr, R&D, #4335-MC-100; PARP1, ABclonal, #A0010; AIF, Santa Cruz Biotechnology, #sc-13116; TOMM20, Beyotime, #AF1717; MIF, Beyotime, #AF7467; β-Tubulin, Beyotime, #AF2839; ERK1, Beyotime, #AF1315; Phospho-Erk1 (Thr202/Tyr204)/Erk2 (Thr185/Tyr187), Beyotime, #AF1891); MEK1/2, Beyotime, #AF1057; Phospho-MEK1 (Ser218/222), Beyotime, #AF1786; DUSP1, MCE, #HY- P80645 ; GAPDH, ABclonal, # AC002) and the secondary antibody (ABclonal, #AS014; ABclonal, #AS003).

Techniques: Cell Culture, Expressing, Western Blot, Membrane, Staining

Journal: Redox Report : Communications in Free Radical Research

Article Title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

doi: 10.1080/13510002.2025.2588086

Figure Lengend Snippet: BS affects NF-κB, p38/JNK, and PI3K-Akt signaling pathways in lung cancer. (a–d) A549 and H1299 cells were treated with BS (0, 5, 10 μM) for 24 h, followed by western blot. Error bars are means ± std. * P < 0.05, ** P < 0.01, *** P < 0.001, compared with the control group by One-way ANOVA (n = 3).

Article Snippet: The primary antibodies used for immunoblotting analysis are as follows: TrxR1 (Proteintech, 11117-1-AP), Trx1 (Proteintech, 14999-1-AP), HBP1 (Proteintech, 11746-1-AP), DNMT1 (Proteintech, 24206-1-AP), Bcl-2 (Proteintech, 12789-1-AP), Bax (Proteintech, 50599-2-Ig), β-actin (Bioss, bs-0061R), Flag (Sigma-Aldrich, F1804), HA (Covance, MMS-101P), p53 (Santa, sc-126), p21 (MBL, K0081-3), p27 (MBL, K0082), γ-H2AX (CST, 9718), NF-κB (Abcam, ab32536), p-NF-κB(CST, 3033), p38 (Santa, sc-7972), p-p38 (Santa, sc-101759), JNK (MCE, HY- P80728 ), p-JNK (Immunoway, YP0157), Akt (Santa, sc-5298), phospho-Akt (CST, 4060).

Techniques: Protein-Protein interactions, Western Blot, Control

Journal: Redox Report : Communications in Free Radical Research

Article Title: The novel thioredoxin reductase inhibitor butaselen suppresses lung cancer by inducing oxidative stress

doi: 10.1080/13510002.2025.2588086

Figure Lengend Snippet: Schematic model of lung cancer inhibition by BS. The TrxR/Trx inhibitor butaselen (BS) can inhibit lung cancer by inducing ROS-dependent apoptosis. The inactivation of the NF-κB and PI3K-Akt signaling pathways, along with the activation of the p38/JNK signaling pathway, contributes to the anti-cancer effects of BS on lung cancer. Although p53 itself is not activated by BS, the HBP1/DNMT1/p21/γ-H2AX/Bcl-2/Bax signaling pathway is activated by BS and contributes to its tumor-inhibitory role. Further mechanistic studies revealed HBP1 as a novel target of the Trx system. The Trx system inversely associates with HBP1 in lung cancer and regulates HBP1 expression at the post-translational level. Under normal conditions, TrxR1 catalyzes the reduction of Trx1 by utilizing NADPH. In its reduced form, Trx1 interacts with HBP1, promoting the ubiquitination of HBP1, which leads to its proteasomal degradation and maintains a low level of HBP1 within cancer cells. Treatment with butaselen inhibits the activity of TrxR1 in lung cancer cells, resulting in the oxidation of Trx1 and the subsequent excessive generation of ROS. HBP1 is activated after being released by the oxidized Trx1 and escaping proteasomal degradation. The activated HBP1 inhibits the expression of DNMT1 and elevates Bax. The decreased DNMT1 further results in the demethylation of the whole genome as well as the promoters of p21 and HOXA9. Ultimately, the upregulation of p21 and γ-H2AX, along with the downregulation of DNMT1 and Bcl-2/Bax, contributes to the apoptosis of lung cancer cells induced by BS. Taken together, the TrxR/Trx inhibitor butaselen inhibits lung cancer by promoting ROS-induced apoptosis through the NF-κB, PI3K-Akt, p38/JNK, and HBP1/DNMT1 signaling pathways.

Article Snippet: The primary antibodies used for immunoblotting analysis are as follows: TrxR1 (Proteintech, 11117-1-AP), Trx1 (Proteintech, 14999-1-AP), HBP1 (Proteintech, 11746-1-AP), DNMT1 (Proteintech, 24206-1-AP), Bcl-2 (Proteintech, 12789-1-AP), Bax (Proteintech, 50599-2-Ig), β-actin (Bioss, bs-0061R), Flag (Sigma-Aldrich, F1804), HA (Covance, MMS-101P), p53 (Santa, sc-126), p21 (MBL, K0081-3), p27 (MBL, K0082), γ-H2AX (CST, 9718), NF-κB (Abcam, ab32536), p-NF-κB(CST, 3033), p38 (Santa, sc-7972), p-p38 (Santa, sc-101759), JNK (MCE, HY- P80728 ), p-JNK (Immunoway, YP0157), Akt (Santa, sc-5298), phospho-Akt (CST, 4060).

Techniques: Inhibition, Protein-Protein interactions, Activation Assay, Expressing, Ubiquitin Proteomics, Activity Assay

Journal: International Journal of Biological Sciences

Article Title: N 6 -methyladenosine Reader IGF2BP2-modified HMMR Promotes Non-small Cell Lung Cancer Metastasis via Interaction with MAP4K4

doi: 10.7150/ijbs.104097

Figure Lengend Snippet: MAP4K4 inhibitor GNE-495 reduces NSCLC cell metastasis in vivo . (A) Flowchart illustrating the NSCLC cell in vivo metastasis model. (B) Relative mRNA and protein expression levels of HMMR and MMP1 in the A549 cells used to establish the in vivo metastasis model. (C) Photographs showing lung metastatic nodules in mice following injection of A549 cells overexpressing HMMR or the control vector, with treatment by DMSO or GNE-495. Metastatic nodules are indicated by red arrowheads. (D) Quantification of lung metastatic nodules across the four experimental groups. (E) The lung weights of the mice in each group after execution. (F) Hematoxylin and eosin (H&E) staining of lung tissue to assess micrometastatic foci. Representative histological images of micrometastatic foci in the four groups. Micrometastatic foci are indicated by blue arrowheads.

Article Snippet: From the fifth week post-injection, mice received daily intraperitoneal injections of either DMSO (3 mg/kg) or the MAP4K4 inhibitor GNE-495 (HY-100343, MCE, China) (3 mg/kg).

Techniques: In Vivo, Expressing, Injection, Control, Plasmid Preparation, Staining

Journal: Pharmaceuticals

Article Title: Identification of Potential JNK3 Inhibitors: A Combined Approach Using Molecular Docking and Deep Learning-Based Virtual Screening

doi: 10.3390/ph16101459

Figure Lengend Snippet: The signal transduction pathways and the hybrid virtual screening process of the JNK3 target.

Article Snippet: The datasets employed in this study consist of two parts: the JNK3 inhibitor activity dataset, containing 1072 molecules which were obtained from the ChEMBL database, is used for training, evaluation, and testing the performance of the rescoring module; the “in-house” database, assembled using the MCE-like compound library and Chemdiv compounds, which contains about 1,600,000 molecules, serves the screening task.

Techniques: Transduction

Journal: Pharmaceuticals

Article Title: Identification of Potential JNK3 Inhibitors: A Combined Approach Using Molecular Docking and Deep Learning-Based Virtual Screening

doi: 10.3390/ph16101459

Figure Lengend Snippet: ( A ) The 3D binding mode of the JNK3 with its original ligand 4-[5-(2-chloro-6-fluoroanilino)-6-methyl-1H-pyrazolo[3,4-b] pyridin-1-yl]-N-(oxetan-3-yl) thiophene-2-carboxamide (PDB ID: 7S1N). ( B ) The binding conformation of the ligand in JNK3 complex. ( C ) The hydrophobicity of the binding pocket. ( D ) The electricity of the binding pocket.

Article Snippet: The datasets employed in this study consist of two parts: the JNK3 inhibitor activity dataset, containing 1072 molecules which were obtained from the ChEMBL database, is used for training, evaluation, and testing the performance of the rescoring module; the “in-house” database, assembled using the MCE-like compound library and Chemdiv compounds, which contains about 1,600,000 molecules, serves the screening task.

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Identification of Potential JNK3 Inhibitors: A Combined Approach Using Molecular Docking and Deep Learning-Based Virtual Screening

doi: 10.3390/ph16101459

Figure Lengend Snippet: ( A ) The inhibition rate (%) assay of 10 selected compounds against JNK3 (Staurosporine is the reference compound); ( B ) the IC 50 evaluation of compound 6; ( C ) TNF-α secretion study. Pos: JNK Inhibitor VIII, Sam: compound; ( D ) IL6 secretion study. ** represents a p -value less than 0.01; *** represents a p -value less than 0.001.

Article Snippet: The datasets employed in this study consist of two parts: the JNK3 inhibitor activity dataset, containing 1072 molecules which were obtained from the ChEMBL database, is used for training, evaluation, and testing the performance of the rescoring module; the “in-house” database, assembled using the MCE-like compound library and Chemdiv compounds, which contains about 1,600,000 molecules, serves the screening task.

Techniques: Inhibition

Journal: Pharmaceuticals

Article Title: Identification of Potential JNK3 Inhibitors: A Combined Approach Using Molecular Docking and Deep Learning-Based Virtual Screening

doi: 10.3390/ph16101459

Figure Lengend Snippet: ( A ) RMSD value of JNK3; ( B ) RMSD value of compound 6; ( C ) the contribution of compound 6-JNK3 complex; ( D ) the 3D diagram of protein–ligand interaction.

Article Snippet: The datasets employed in this study consist of two parts: the JNK3 inhibitor activity dataset, containing 1072 molecules which were obtained from the ChEMBL database, is used for training, evaluation, and testing the performance of the rescoring module; the “in-house” database, assembled using the MCE-like compound library and Chemdiv compounds, which contains about 1,600,000 molecules, serves the screening task.

Techniques: