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: The Journal of endocrinology

Article Title: The regulatory mechanism by which interleukin-6 stimulates GH-gene expression in rat GH3 cells.

doi: 10.1677/joe.1.06736

Figure Lengend Snippet: Figure 4 Effects of IL-6 on MEK and p38 MAPK (total and phosphorylated) activities in GH3 cells. Stably transfected GH3 cells were stimulated with 103 U/ml (250 ng/ml) IL-6 in serum-free medium for the indicated time (at top of a and c) in the presence or absence of 40 mM PD98059 (a and b) or 5 mM SB203580 (c and d). Cells were then lysed in SDS sample buffer and proteins were separated by electrophoresis with subsequent electrotransferring to nitrocellulose membrane. Immunoblotting detection was performed using (a) anti-MEK, (b) anti-phospho-specific MEK, (c) antibodies or anti-p38 MAPK, and (d) anti-phospho-specific p38 MAPK antibodies as the primary antibodies at a 1:1000 dilution. The positions of total MEK (t-MEK), phosphorylated MEK (p-MEK), total MAPK (t-p38 MAPK), and phosphorylated MAPK (p-p38 MAPK) are indicated on the right-hand side of each panel. On the left-hand side is the protein molecular marker (kDa). The blots are representative of three independent experiments.

Article Snippet: Protein extraction, electrophoresis, and Western blotting Protein extraction, gel electrophoresis, and Western blot analysis were performed according to the manufacturer’s instructions of the commercially available PhosphoPlus MEK1/2 and p38 MAP Kinase antibody kits (Cell Signaling Technology, Beverly, MA, USA).

Techniques: Stable Transfection, Transfection, Electrophoresis, Membrane, Western Blot, Marker

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Hawthorne leaf flavonoids prevent oxidative stress injury of renal tissues in rats with diabetic kidney disease by regulating the p38 MAPK signaling pathway

doi:

Figure Lengend Snippet: Immunochemical analysis of the (A) p38MAPK (B) and p-p38MAPK expression in the kidney of different groups (200×). Quantitative analysis of (C) p38MAPK and (D) p-p38MAPK expression in the kidney of different groups. The expression of p38MPAK and p-p38MAPK was significantly higher in the DKD group than in the HLF and IRB groups.

Article Snippet: Rabbit anti-rat p38MAPK and p-p38MAPK polyclonal antibodies were purchased from Boster (Beijing, China).

Techniques: Expressing

Journal: Molecular medicine reports

Article Title: Placental growth factor gene silencing mitigates the epithelial‑to‑mesenchymal transition via the p38 MAPK pathway in rats with hyperoxia‑induced lung injury.

doi: 10.3892/mmr.2019.10785

Figure Lengend Snippet: Figure 5. Expression of p38 and p‑p38 MAPK in hyperoxia‑exposed lung tissue. Neonatal rats were exposed to normoxic or hyperoxic conditions for 14 days; hyperoxia exposed rats were subsequently injected with shRNA‑NC or shRNA‑PLGF. (A) Western blot analysis and (B) quantification was performed to determine p‑p38/p38 protein expression levels in rat lung tissues; β‑actin was used as the endogenous control. Data are presented as the mean ± standard deviation. n=8. ***P<0.001. MAPK, mitogen‑activated protein kinase; NC, negative control; PLGF, placental growth factor; shRNA, short hairpin RNA.

Article Snippet: The animals were housed at a temperature of 25‐27 ̊C, with a humidity of 50-70% and a 12 h light/dark cycle with ad libitum access to food and water. xylene, absolute ethanol, eosin y and hydrogen peroxide were purchased from Wuhan uScn Business co., ltd. Hematoxylin, eosin and goat serum (cat. no. Sl038) were purchased from Beijing Solarbio Science& Technology co., ltd. PlGF mouse monoclonal antibody (cat. no. sc-518003) and e-cadherin mouse monoclonal antibody (cat. no. sc-71007) were purchased from Santa cruz Biotechnology, inc. anti-phosphorylated (p)-p38 rabbit polyclonal antibody (cat. no. bs-2210r) was purchased from (BioSS). anti-p38 rabbit monoclonal (cat. no. M00176), anti-β-actin goat polyclonal (cat. no. BM0627) and anti-ZeB2 rabbit polyclonal (cat. no. Pa1959) antibodies were purchased from Boster Biological Technology.

Techniques: Expressing, Injection, Western Blot, Control, Standard Deviation, Negative Control, shRNA

Journal: Frontiers in pharmacology

Article Title: Neuroprotective Effects and Mechanisms of Zhenlong Xingnao Capsule in In Vivo and In Vitro Models of Hypoxia.

doi: 10.3389/fphar.2019.01096

Figure Lengend Snippet: FIGURE 6 | Effects of ZXC on the mRNA levels of the Bcl-2/Bax ratio (A), caspase-3 (B), nuclear factor (NF)-кB (C), and p38 (D) in the prefrontal cortex of ischemia-reperfusion injury rats. The data are expressed as mean ± standard deviation (n = 3). I-30, ischemia for 30 min; I-90, ischemia for 90 min; I-90+R-30, ischemia for 90 min, then reperfusion for 30 min; I-90+R-180, ischemia for 90 min, then reperfusion for 180 min. *P < 0.05 vs. sham group; #P < 0.05 vs. model group; ##P < 0.01 vs. model group.

Article Snippet: The primary antibodies used for IHC were the MAPK14 (p38) antibody and CASP3 (P17) antibody; both were obtained from Boster Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Standard Deviation

Journal: Frontiers in pharmacology

Article Title: Neuroprotective Effects and Mechanisms of Zhenlong Xingnao Capsule in In Vivo and In Vitro Models of Hypoxia.

doi: 10.3389/fphar.2019.01096

Figure Lengend Snippet: FIGURE 7 | Western blotting results for Sham, Model and ZXC groups (A). Effects of ZXC on the protein expressions Bcl-2 (B), Bax (C), Caspase-3 (D), NF-кB (E), and p38 (F) in brain tissue of ischemia-reperfusion injury rats induced by MCAO. The data are expressed as mean ± standard deviation (n = 4). I-30, ischemia for 30 min; I-90, ischemia for 90 min; I-90+R-30, ischemia for 90 min, then reperfusion for 30 min. *P < 0.05 vs. sham group; #P < 0.05 vs. model group.

Article Snippet: The primary antibodies used for IHC were the MAPK14 (p38) antibody and CASP3 (P17) antibody; both were obtained from Boster Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Western Blot, Standard Deviation

Journal: Frontiers in Pharmacology

Article Title: Computational and In Vitro Analysis of Plumbagin’s Molecular Mechanism for the Treatment of Hepatocellular Carcinoma

doi: 10.3389/fphar.2021.594833

Figure Lengend Snippet: (A) Protein expression levels of LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK in SMMC-7721 cells measured by Western blot after treatment with 5 μM PL, 10 μM 3-MA autophagy inhibitor, 10 μM rapamycin autophagy agonist, 10 μM SB202190, and 10 μM SB203580 p38 MAPK inhibitor. (B) LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK were used as internal reference total proteins for gray value analysis. * p < 0.05, ** p < 0.01 *** p < 0.001, compared with the PL group. # p < 0.05, ## p < 0.01 ### p < 0.001 vs. the control group. (C) Protein expression levels of LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK in BEL-7404 cells measured by Western blot after treatment with 5 μM PL, 10 μM 3-MA autophagy inhibitor, 10 μM rapamycin autophagy agonist, 10 μM SB202190, and 10 μM SB203580 p38 MAPK inhibitor. (D) LC3-I, LC3-II, p38 MAPK, and p-p38 MAPK were used as internal reference total proteins for gray value analysis. * p < 0.05, ** p < 0.01 *** p < 0.001, compared with the PL group. # p < 0.05, ## p < 0.01 ### p < 0.001 vs. the control group.

Article Snippet: Plumbagin (PL) was purchased from Sigma-Aldrich (St. Louis, MO, United States) with purity ≥98%; the Acridine Orange (AO)/Ethidium bromide (EB) Double Stain Kit was from Solable Technology (Beijing, China); N-acetyl-l-cysteine, SB203580, and SB202190 were from Sigma-Aldrich (St. Louis, MO, United States); SC-79, MEK2206, 3-MA, and Z-VAD-FMK were from Selleck (Texas, United States); the BCA Protein Assay Kit, ROS Assay Kit, Annexin V-FITC Apoptosis Detection Kit and Cell lysis buffer for Western were all obtained from Beyotime Biotechnology (Shanghai, China); antibodies against Akt, phospho-Akt, mTOR, phospho-mTOR, p38 MAPK, phospho-p38 MAPK, PI3K, phospho-PI3K, LC3B, cleave-RP, and cleave-caspase 3 were from Cell Signaling Technology, Inc. (Boston, MA, United States); and β-p38 MAPK was purchased from Boster Technology (Wuhan, China).

Techniques: Expressing, Western Blot, Control

Journal: NPJ Science of Food

Article Title: Effects of Mytilus edulis derived plasmalogens against atherosclerosis via lipid metabolism and MAPK signaling pathway

doi: 10.1038/s41538-025-00546-0

Figure Lengend Snippet: HFD has a significant pro-atherogenic effect, which might be associated with provocation of AP-1 mediated inflammatory response mediated by calmodulin. In contrast, Pls supplementation effectively inhibits HFD-induced activation of MAPK signaling pathway, thereby preventing and the downstream inflammatory damage and blocking the development of HFD-related disorders, including atherosclerosis.

Article Snippet: Subsequently, the membranes were washed with TBST three times, and incubated with the following primary antibodies: β-actin (HY- P80438 , MedChemExpress, Monmouth Junction, NJ, USA), p38 MAPK (HY- P80776 , MedChemExpress), Casp1 (ab138483, abcam, Cambridge, UK), Calm4 (ab2860, abcam), and Tnf (HY-P7090, MedChemExpress) at 4 °C overnight with agitation, followed by incubation with the second antibody for 4 h. The protein bands were visualized using electrochemiluminescence, and the intensity of bands was quantitatively analyzed using ImageJ software, with β-actin as the protein loading control.

Techniques: Activation Assay, Blocking Assay

Journal: Materials

Article Title: A Novel Bioactive Endodontic Sealer Containing Surface-Reaction-Type Prereacted Glass-Ionomer Filler Induces Osteoblast Differentiation

doi: 10.3390/ma13204477

Figure Lengend Snippet: ( A ) mRNA expression of ALP and IBSP induced by set PRG+ extract significantly downregulated by application of NPS2143 (CaSR antagonist) in Kusa-A1 cells; * p < 0.05. ( B ) Phosphorylation of extracellular signal-regulated kinase (ERK) and p38 promoted by set PRG+ extract is downregulated by the application of NPS2143. Phosphorylation of ERK and p38 promoted by Sr 2+ is also downregulated by NPS2143 application. α-Tubulin was used as loading control. Experiments were repeated three times with similar results.

Article Snippet: Membranes were incubated with 1:1000-diluted polyclonal antibodies against extracellular signal-regulated kinase (ERK), phosphorylated ERK (pERK; 1:1000; BD Bioscience, San Jose, CA USA), p38 mitogen-activated protein kinase (MAPK), and phosphorylated p38 MAPK (pp38; BD Bioscience) overnight at 4 °C.

Techniques: Expressing