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: 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: 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: 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

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: BMC Oral Health

Article Title: Artesunate-loaded thermosensitive chitosan hydrogel promotes osteogenesis of maxillary tooth extraction through regulating T lymphocytes in type 2 diabetic rats

doi: 10.1186/s12903-024-04127-7

Figure Lengend Snippet: ART-loaded TCH suppressed MAPK signaling pathway in the maxilla of T2DM tooth extraction. A Network pharmacology analysis revealed the potential immunoregulatory mechanism of ART in T2DM. The top ten enriched pathways were illustrated. B Molecular docking of ART to MAPK and ERK1 proteins. C, D The protein levels of p38-MAPK, p-p38-MAPK, ERK1, and p-ERK1, were analyzed by WB and IHC ( n = 3), which can be downregulated by ART-loaded TCH treating. The uncropped blots are displayed in Additional file . E Relative mRNA expression of MAPK ( D ) and ERK1 ( E ) was analyzed by PCR ( n = 3). The 150 mg/ml ART-loaded TCH group significantly decreased the mRNA expression of MAPK and ERK1 than the T2DM group ( p < 0.01). Data are shown as mean ± standard deviation. * p < 0.05 and ** p < 0.01. T2DM, type 2 diabetes mellitus group; TCH, thermosensitive chitosan hydrogel group; Low, 50 mg/ml ART-loaded TCH group; Medium, 100 mg/ml ART-loaded TCH group; High; 150 mg/ml ART-loaded TCH group

Article Snippet: The membranes were then incubated overnight with primary antibodies including rabbit anti-β-actin (1:20000, Bioss), mouse anti-GATA-3 (1:2000, Proteintech), mouse anti-IL-4 (1:1000, Proteintech), rabbit anti-T-BET (1:500, Proteintech), rabbit anti-phospho-p38 MAPK (1:1000, Bioss), rabbit anti-p38 MAPK (1:1000, Bioss), mouse anti-ERK1 (1:1000, Bioss), mouse anti-phospho-ERK1 (1:1000, Bioss).

Techniques: Extraction, Expressing, Standard Deviation

Journal: eLife

Article Title: Domain fusion TLR2-4 enhances the autophagy-dependent clearance of Staphylococcus aureus in the genetic engineering goat

doi: 10.7554/eLife.78044

Figure Lengend Snippet: ( A ) The change in phosphorylation of JNK, ERK, and p38 in S. aureus -infected macrophages. Macrophages were pretreated with DMSO (carrier), Takinib (200 μM), or Amlexanox (100 μM) for 12 hr, then stimulated with S. aureus for 4 hr. Phosphorylation of JNK, ERK, and p38 was observed by Western blot. ( B ) The level of LC3-II was detected in macrophages after S. aureus challenge. Macrophages were pretreated with DMSO, Dynasore (50 μM), SP600125 (200 μM), PD98059 (200 μM), SB203580 (200 μM), or Amlexanox for 12 hr, respectively, then treated with S. aureus (multiplicity of infection [MOI] = 10) for 4 hr. Conversion of LC3-I to LC3-II was monitored by Western blot. GAPDH protein was used as a control. ( C ) The effect activating JNK and ERK1/2 on removal of S. aureus in macrophages. Macrophages were treated with DMSO, SP600125, or PD98059 for 12 hr prior to stimulation with fluorescein isothiocyanate (FITC)-labeled S. aureus (MOI = 10) for 4 hr. The mean fluorescence intensity (MFI) of FITC in macrophages was measured by flow cytometry. ( D ) The expression of TFEB and OPTN was analyzed in macrophages by Western blot. Macrophages were pretreated with DMSO, Dynasore, or Amlexanox for 12 hr, then challenged with S. aureus for 4 hr (MOI = 10). The nucleoprotein was extracted and analyzed by Western blotting to detect the translocation of TFEB. The expression of OPTN was detected by Western blot. LMNB2 and GAPDH were used to normalize samples. Figure 5—source data 1. The original blots of . Figure 5—source data 2. The original blots of . Figure 5—source data 3. The original blots of .

Article Snippet: Antibody , Phospho-p38 MAPK-T180 (Rabbit polyclonal) , ABclonal , Cat#: AP0238 RRID: AB_2771307 , WB (1:1000).

Techniques: Phospho-proteomics, Infection, Western Blot, Control, Labeling, Fluorescence, Flow Cytometry, Expressing, Translocation Assay

Journal: eLife

Article Title: Domain fusion TLR2-4 enhances the autophagy-dependent clearance of Staphylococcus aureus in the genetic engineering goat

doi: 10.7554/eLife.78044

Figure Lengend Snippet:

Article Snippet: Antibody , Phospho-p38 MAPK-T180 (Rabbit polyclonal) , ABclonal , Cat#: AP0238 RRID: AB_2771307 , WB (1:1000).

Techniques: Bacteria, Transfection, Construct, Sequencing, Control, Extraction, Software, Staining

Journal: Toxics

Article Title: The Role of G3BP1 Gene Mediates P38 MAPK/JNK Pathway in Testicular Spermatogenic Dysfunction Caused by Cyfluthrin

doi: 10.3390/toxics11050451

Figure Lengend Snippet: Expression changes of key factors of the MAPK pathway in testicular tissue detected by q-PCR. ( A – F ) The relative mRNA expression of JNK1 , JNK2 , JNK3 , ERK1 , ERK2 and P38MAPK. Compared to the control group, * p < 0.05 and ** p < 0.01.

Article Snippet: Phosphp-ERK1/2 antibody, phosphp-JNK1/2/3 antibody, phosphp-p38MAPK antibody, rabbit anti-caspase 3 polyclonal antibody, rabbit anti-caspase 8 polyclonal antibody, rabbit anti-caspase 9 polyclonal antibody, rabbit anti-GAPDH polyclonal antibody, goat anti-rabbit IgG (H + L) HRP and goat anti-mouse IgG-HRP were provided by Affinity Biosciences Ltd. (Liyang, China).

Techniques: Expressing