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 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: Infection and Immunity

Article Title: Thioredoxin from Brugia malayi: Defining a 16-Kilodalton Class of Thioredoxins from Nematodes

doi: 10.1128/IAI.71.7.4119-4126.2003

Figure Lengend Snippet: Bm-TRX-1 inhibits activation of p38 MAP kinase. The WCPPCR and WCPQCR alleles of Bm-trx-1 were cloned separately into a eukaryotic expression plasmid vector, and the plasmids were introduced into TPA-differentiated ML-1 cells. The transfected ML-1 cells were stimulated with TNF-α (100 IU/well) for 5 min, and the phosphorylation status of p38 MAP kinase was analyzed on immunostained Western blots of cell extracts. Controls included cells maintained in medium (medium control), cells exposed to the transfection regent (transfection control), and cells transfected with a plasmid containing no insert (plasmid control).

Article Snippet: In preliminary experiments, TPA-differentiated ML-1 cells were stimulated with TNF-α (100 IU/well) for 5, 10, and 30 min, and the phosphorylation status of p38 MAP kinase was analyzed on immunostained Western blots of cell extracts with phosphoPlus p38 MAP kinase antibody kit (Cell Signaling Technology, Beverly, Mass.).

Techniques: Activation Assay, Clone Assay, Expressing, Plasmid Preparation, Transfection, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: MMP28 recruits M2-type tumor-associated macrophages through MAPK/JNK signaling pathway-dependent cytokine secretion to promote the malignant progression of pancreatic cancer

doi: 10.1186/s13046-025-03321-x

Figure Lengend Snippet: MMP28 promotes the secretion of IL-8 and VEGFA from cancer cells by regulating the MAPK/JNK signaling pathway. A - B WB analysis of JNK, p-JNK, ERK, p-ERK, P38, and p-P38 in pancreatic cancer cells with MMP28 knockdown or overexpression compared with the control. C - D GEPIA database analysis of the relationships between JNK expression and the expression of IL-8 and VEGFA in pancreatic cancer. E ELISAs were used to measure IL-8 and VEGFA protein secretion in empty vector-treated pancreatic cancer cells, MMP28-overexpressing pancreatic cancer cells, and pancreatic cancer cells treated with the JNK inhibitor SP600125 (20 μM). **** P < 0.0001. ### P < 0.01, and #### P < 0.0001

Article Snippet: The membranes were blocked with 5% bovine serum albumin (GC305010-5 g, Servicebio) for one hour and incubated overnight at 4°C with primary antibodies against MMP28 (1:500, sc-515010, Santa Cruz Biotechnology), GAPDH (1:1000, GB12002, Servicebio), β-actin (1:1000, GB15003-100, Servicebio), JNK (1:500, BM4329, BOSTER), p-JNK (1:500 BM4380, BOSTER), P38-MAPK (1:500, BM4439, BOSTER), p-P38-MAPK (1:500, M00176T180, BOSTER), ERK (1:500, BM3426, BOSTER), p-ERK (1:500, BM4156, BOSTER), ANXA2 (1:1000, 11256-1-AP, Proteintech), Vimentin (1:500, PB9359, BOSTER), CDH1 (1:500, BM4166, BOSTER), CDH2 (1:500, BA0673, BOSTER), BAX (1:500, BA0315-2, BOSTER), and BCL-2 (1:500, A00040-2, BOSTER).

Techniques: Knockdown, Over Expression, Control, Expressing, Plasmid Preparation

Journal: Cell Biology and Toxicology

Article Title: TEAD4-mediated upregulation of LPAR3 augments hepatic stellate cell activation in portal hypertension

doi: 10.1007/s10565-025-10063-1

Figure Lengend Snippet: The LPAR3/p38 MAPK/PI3K/AKT cascades are activated in TGF-β1-stimulated HSCs. LX-2 cells were treated with treated with 10 ng/mL recombinant human TGF-β1 or PBS. A , expression of α-SMA in cells determined by cell immunofluorescence staining; B , LPAR3 mRNA expression in cells determined using RT-qPCR; C - G , protein concentration of LPAR3, and phosphorylation levels of p38 MAPK, PI3K, and AKT in LX-2 cells determined by WB analysis. Three independent experiments were performed. Differences were analyzed by the unpaired t-test

Article Snippet: After blocking, the sections were probed with the Myosin IIA antibody (1:500, ab238131, Abcam) at 22–25°C for 2 h, followed by incubation with IgG (Alexa Fluor 488) (1:1000, ab150077, Abcam) at 22–25°C in darkness for 1 h. Additionally, fluorescent primary antibodies were added as needed and incubated in the dark for 1 h. These antibodies included AbBy Fluor 488-conjugated anti-LPAR3 (1:200, bs-2882R-BF488, Bioss Antibodies, Woburn, MA, USA), anti-phospho-p38 MAPK (1:200, bs-23251R-BF488, Bioss), anti-phospho-PI3K (1:200, bs-5571R-BF488, Bioss), anti-phospho-AKT (1:200, bs-12456R-BF488, Bioss), anti-TEAD4 (1:200, bs-9603R-BF488, Bioss), and Alexa Fluor 647-conjugated anti-α-SMA (1:200, ITT5797-647, G-Biosciences, St Louis, MO, USA).

Techniques: Recombinant, Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Protein Concentration, Phospho-proteomics

Journal: Cell Biology and Toxicology

Article Title: TEAD4-mediated upregulation of LPAR3 augments hepatic stellate cell activation in portal hypertension

doi: 10.1007/s10565-025-10063-1

Figure Lengend Snippet: LPAR3 modulates LX-2 cell activation through the p38 MAPK and PI3K/AKT signaling pathways. LX-2 cells were administered three lentiviral vector-carried shRNAs (shLPAR3 #1, #2, and #3). A , relative mRNA expression of LPAR3 in cells determined using RT-qPCR. LX-2 stably transfected with shLPAR3 were subjected to 10 ng/mL recombinant human TGF-β1 treatment and the additional 100 nM CrPic, 20 μM 740 Y-P, or DMSO. B - F , protein concentration of LPAR3, and phosphorylation levels of p38 MAPK, PI3K, and AKT in LX-2 cells determined by WB analysis; G , contractile activity of HSCs evaluated by the collagen gel contraction assay; H , F-actin expression in LX-2 cells determined using actin tracking-based immunofluorescence staining; I, Collagen I expression in LX-2 cells determined using immunofluorescence staining. Three independent experiments were performed. Differences were analyzed by one-way ANOVA followed by Tukey’s post-hoc test

Article Snippet: After blocking, the sections were probed with the Myosin IIA antibody (1:500, ab238131, Abcam) at 22–25°C for 2 h, followed by incubation with IgG (Alexa Fluor 488) (1:1000, ab150077, Abcam) at 22–25°C in darkness for 1 h. Additionally, fluorescent primary antibodies were added as needed and incubated in the dark for 1 h. These antibodies included AbBy Fluor 488-conjugated anti-LPAR3 (1:200, bs-2882R-BF488, Bioss Antibodies, Woburn, MA, USA), anti-phospho-p38 MAPK (1:200, bs-23251R-BF488, Bioss), anti-phospho-PI3K (1:200, bs-5571R-BF488, Bioss), anti-phospho-AKT (1:200, bs-12456R-BF488, Bioss), anti-TEAD4 (1:200, bs-9603R-BF488, Bioss), and Alexa Fluor 647-conjugated anti-α-SMA (1:200, ITT5797-647, G-Biosciences, St Louis, MO, USA).

Techniques: Activation Assay, Protein-Protein interactions, Plasmid Preparation, Expressing, Quantitative RT-PCR, Stable Transfection, Transfection, Recombinant, Protein Concentration, Phospho-proteomics, Activity Assay, Collagen Gel Contraction Assay, Immunofluorescence, Staining

Journal: Cell Biology and Toxicology

Article Title: TEAD4-mediated upregulation of LPAR3 augments hepatic stellate cell activation in portal hypertension

doi: 10.1007/s10565-025-10063-1

Figure Lengend Snippet: LPAR3 knockdown reduces activation of p38 MAPK and PI3K/AKT signaling and alleviates HSC activation in mice with PHT. A mouse model of cirrhotic PHT was generated by injection of a mixture of 5 mL/kg CCl 4 , with olive oil (vehicle) injection alone serving as control. Lentiviral vector-carried shLPAR3 or shScramble was administered during the first CCl 4 injection. A , a schematic diagram for animal treatment; B , portal pressure of mice after week 6; C-D , positive staining area of α-SMA and LPAR3 in the mouse liver tissue sections examined using double-label IHC assay; E , mean fluorescence intensities of p-p38 MAPK, p-PI3K, and p-AKT in HSCs (α-SMA positive) analyzed using double-label IHC assays, with 50 HSCs from each section included for assessment. Each group contained 6 mice. Differences were analyzed by one-way ANOVA followed by Tukey's post-hoc test

Article Snippet: After blocking, the sections were probed with the Myosin IIA antibody (1:500, ab238131, Abcam) at 22–25°C for 2 h, followed by incubation with IgG (Alexa Fluor 488) (1:1000, ab150077, Abcam) at 22–25°C in darkness for 1 h. Additionally, fluorescent primary antibodies were added as needed and incubated in the dark for 1 h. These antibodies included AbBy Fluor 488-conjugated anti-LPAR3 (1:200, bs-2882R-BF488, Bioss Antibodies, Woburn, MA, USA), anti-phospho-p38 MAPK (1:200, bs-23251R-BF488, Bioss), anti-phospho-PI3K (1:200, bs-5571R-BF488, Bioss), anti-phospho-AKT (1:200, bs-12456R-BF488, Bioss), anti-TEAD4 (1:200, bs-9603R-BF488, Bioss), and Alexa Fluor 647-conjugated anti-α-SMA (1:200, ITT5797-647, G-Biosciences, St Louis, MO, USA).

Techniques: Knockdown, Activation Assay, Generated, Injection, Control, Plasmid Preparation, Staining, Fluorescence

Journal: Cell Biology and Toxicology

Article Title: TEAD4-mediated upregulation of LPAR3 augments hepatic stellate cell activation in portal hypertension

doi: 10.1007/s10565-025-10063-1

Figure Lengend Snippet: Silencing of TEAD4 alleviates activation and contraction of LX-2 cells, which are negated by LPAR3 restoration. LX-2 cells were administered three lentiviral vector-carried shRNAs (sh TEAD4 #1, #2, and #3). A , relative mRNA expression of LPAR3 in cells determined using RT-qPCR. LX-2 stably transfected with shTEAD4 were additionally administered Vector-LPAR3, followed by 10 ng/mL recombinant human TGF-β1 treatment. B - G , protein concentration of TEAD4 and LPAR3, and phosphorylation levels of p38 MAPK, PI3K, and AKT in LX-2 cells determined by WB analysis; H , contractile activity of HSCs evaluated by the collagen gel contraction assay; I , α-SMA expression in LX-2 cells determined using immunofluorescence assay. Three independent experiments were performed. Differences were analyzed by one-way ANOVA followed by Tukey's post-hoc test

Article Snippet: After blocking, the sections were probed with the Myosin IIA antibody (1:500, ab238131, Abcam) at 22–25°C for 2 h, followed by incubation with IgG (Alexa Fluor 488) (1:1000, ab150077, Abcam) at 22–25°C in darkness for 1 h. Additionally, fluorescent primary antibodies were added as needed and incubated in the dark for 1 h. These antibodies included AbBy Fluor 488-conjugated anti-LPAR3 (1:200, bs-2882R-BF488, Bioss Antibodies, Woburn, MA, USA), anti-phospho-p38 MAPK (1:200, bs-23251R-BF488, Bioss), anti-phospho-PI3K (1:200, bs-5571R-BF488, Bioss), anti-phospho-AKT (1:200, bs-12456R-BF488, Bioss), anti-TEAD4 (1:200, bs-9603R-BF488, Bioss), and Alexa Fluor 647-conjugated anti-α-SMA (1:200, ITT5797-647, G-Biosciences, St Louis, MO, USA).

Techniques: Activation Assay, Plasmid Preparation, Expressing, Quantitative RT-PCR, Stable Transfection, Transfection, Recombinant, Protein Concentration, Phospho-proteomics, Activity Assay, Collagen Gel Contraction Assay, Immunofluorescence

Journal: Cell Biology and Toxicology

Article Title: TEAD4-mediated upregulation of LPAR3 augments hepatic stellate cell activation in portal hypertension

doi: 10.1007/s10565-025-10063-1

Figure Lengend Snippet: LPAR3 upregulation augments liver injury in PHT mice alleviated by TEAD4 silencing. A mouse model of cirrhotic PHT was generated by injection of a mixture of 5 mL/kg CCl 4 . Lentiviral vector-carried shTEAD4/shScramble, or the additional Vector-LPAR3/Vector-NC was administered during the first CCl 4 injection. A , portal pressure of mice after week 6; B , positive staining area of α-SMA in mouse liver tissue sections, as well as the mean fluorescence intensity of TEAD4 in 50 HSCs, examined using double-label IHC assay; C - F , mean fluorescence intensities of p-p38 MAPK, p-PI3K, and p-AKT in HSCs (α-SMA positive) analyzed using double-label IHC assays, with 50 HSCs from each section included for assessment; G , pathological changes in mice determined using HE staining. Each group contained 6 mice. Differences were analyzed by one-way ANOVA followed by Tukey's post-hoc test

Article Snippet: After blocking, the sections were probed with the Myosin IIA antibody (1:500, ab238131, Abcam) at 22–25°C for 2 h, followed by incubation with IgG (Alexa Fluor 488) (1:1000, ab150077, Abcam) at 22–25°C in darkness for 1 h. Additionally, fluorescent primary antibodies were added as needed and incubated in the dark for 1 h. These antibodies included AbBy Fluor 488-conjugated anti-LPAR3 (1:200, bs-2882R-BF488, Bioss Antibodies, Woburn, MA, USA), anti-phospho-p38 MAPK (1:200, bs-23251R-BF488, Bioss), anti-phospho-PI3K (1:200, bs-5571R-BF488, Bioss), anti-phospho-AKT (1:200, bs-12456R-BF488, Bioss), anti-TEAD4 (1:200, bs-9603R-BF488, Bioss), and Alexa Fluor 647-conjugated anti-α-SMA (1:200, ITT5797-647, G-Biosciences, St Louis, MO, USA).

Techniques: Generated, Injection, Plasmid Preparation, Staining, Fluorescence

Journal: Cell Biology and Toxicology

Article Title: TEAD4-mediated upregulation of LPAR3 augments hepatic stellate cell activation in portal hypertension

doi: 10.1007/s10565-025-10063-1

Figure Lengend Snippet: Graphical abstract. In the context of PHT, TEAD4 is upregulated, which activates the transcription of LPAR3 and the subsequent p38 MAPK and PI3K/AKT signaling pathways, leading to increased HSC activation and fibrogenesis

Article Snippet: After blocking, the sections were probed with the Myosin IIA antibody (1:500, ab238131, Abcam) at 22–25°C for 2 h, followed by incubation with IgG (Alexa Fluor 488) (1:1000, ab150077, Abcam) at 22–25°C in darkness for 1 h. Additionally, fluorescent primary antibodies were added as needed and incubated in the dark for 1 h. These antibodies included AbBy Fluor 488-conjugated anti-LPAR3 (1:200, bs-2882R-BF488, Bioss Antibodies, Woburn, MA, USA), anti-phospho-p38 MAPK (1:200, bs-23251R-BF488, Bioss), anti-phospho-PI3K (1:200, bs-5571R-BF488, Bioss), anti-phospho-AKT (1:200, bs-12456R-BF488, Bioss), anti-TEAD4 (1:200, bs-9603R-BF488, Bioss), and Alexa Fluor 647-conjugated anti-α-SMA (1:200, ITT5797-647, G-Biosciences, St Louis, MO, USA).

Techniques: Protein-Protein interactions, Activation Assay

Journal: Oncology Letters

Article Title: Tanshinol upregulates the expression of aquaporin 5 in lung tissue of rats with sepsis

doi: 10.3892/ol.2018.9026

Figure Lengend Snippet: Effects of tanshinol on the expression of AQP-5, P38 and p-P38 in lung tissue of rats with Sepsis by western blot analysis. (A) Western blot and quantification of the expression levels of AQP-5 in rats of different group. (B) Western blot and quantification of the levels of P38 and p-P38 in rats of different group. All the data are presented as mean ± standard deviation. **P<0.01 vs. ctrl group; ^^P<0.01 vs. SO group; #P<0.05 vs. Sepsis group; ##P<0.01 vs. Sepsis group. AQP-5, Aquaporin 5; P-, phosphorylated; SO, sham operation group; Sepsis, model group; Tan-L, low dose tanshinol group (5 mg/kg); Tan-M, moderate dose tanshinol group (10 mg/kg); Tan-H, high dose tanshinol group (20 mg/kg). n=20 for all groups.

Article Snippet: The primary antibodies against AQP-5 (1:1,200, catalog no. orb48020), p-P38 (1:1,500, catalog no. orb128324), P38 (1:12,00, catalog no. orb338949) and β-actin (1:2,000, catalog no. orb178392) (all Biorbyt Ltd., Cambridge, UK) were diluted with TBST solution containing 3% bovine serum albumin (catalog no. orb334844, Biorbyt Ltd., Cambridge, UK).

Techniques: Expressing, Western Blot, Standard Deviation