Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Exosomes derived from miR-26a-5p-modified adipose mesenchymal stem cells improve wound healing by targeting MAP2K4

doi: 10.3389/fbioe.2025.1662095

Figure Lengend Snippet: Interaction between miR-26a-5p and MAP2K4 , as well as cell transfection efficiency and characterization of the isolated exosomes. (A) MAP2K4 was the target of miR-26a-5p by dual luciferase reporter gene assay. N = 3. *: P < 0.05. (B) The miR-26a-5p level in the AMSCs with miR-26a-5p overexpression to verify the cell transfection efficiency. N = 3. *: P < 0.05, vs. AMSCs-NC. (C) Particle size distribution of AMSCs-derived exosomes and miR-26a-5p overexpressed AMSCs-derived exosomes determined by Nanosight. (D) The morphology of AMSCs-derived exosomes and miR-26a-5p overexpressed AMSCs-derived exosomes, visualized by transmission electron microscopy. (E) The expression of exosomes-specific markers (CD63, CD81, and HSP70) and negative control protein (calnexin) in exosomes and cells, detected by Western blot. (F) The level of miR-26a-5p in the AMSCs-derived exosomes and miR-26a-5p overexpressed AMSCs-derived exosomes. N = 3. *: P < 0.05, vs. AMSCs-Exo.

Article Snippet: After blocking with 5% skim milk at 37 °C for 2 h, the membranes were incubated the primary antibodies, including anti-CD63 antibody (1: 1,000, Abclonal, Wuhan, China), anti-CD81 antibody (1: 1,000, Abclonal), anti-HSP70 antibody (1: 1,000, Proteintech, Wuhan, China), anti-calnexin antibody (1: 1,000, Proteintech), anti-MAP2K4 antibody (1: 1,000, Proteintech), anti-COL1A1 antibody (1: 1,000, NOVUS, St. Louis, MO, United States), anti-α-SMA antibody (1: 1,000, Cell Signaling Technology, Boston, MA, United States), anti-TNF-α antibody (1: 1,000, Cell Signaling Technology), and anti-GAPDH antibody (1: 10,000, Proteintech).

Techniques: Transfection, Isolation, Luciferase, Reporter Gene Assay, Over Expression, Derivative Assay, Transmission Assay, Electron Microscopy, Expressing, Negative Control, Western Blot

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Exosomes derived from miR-26a-5p-modified adipose mesenchymal stem cells improve wound healing by targeting MAP2K4

doi: 10.3389/fbioe.2025.1662095

Figure Lengend Snippet: The mRNA expression of related genes, including Map2k4 , Col1a1 , Col2a1 , Col3a1 , α-Sma , Tnf-α , Il1β , Il6 , and Cd31 in the different groups measured by RT-qPCR. N = 3. *: P < 0.05, vs. control; # : P < 0.05, vs. model; $ : vs. AMSCs-agomir-Exo.

Article Snippet: After blocking with 5% skim milk at 37 °C for 2 h, the membranes were incubated the primary antibodies, including anti-CD63 antibody (1: 1,000, Abclonal, Wuhan, China), anti-CD81 antibody (1: 1,000, Abclonal), anti-HSP70 antibody (1: 1,000, Proteintech, Wuhan, China), anti-calnexin antibody (1: 1,000, Proteintech), anti-MAP2K4 antibody (1: 1,000, Proteintech), anti-COL1A1 antibody (1: 1,000, NOVUS, St. Louis, MO, United States), anti-α-SMA antibody (1: 1,000, Cell Signaling Technology, Boston, MA, United States), anti-TNF-α antibody (1: 1,000, Cell Signaling Technology), and anti-GAPDH antibody (1: 10,000, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Exosomes derived from miR-26a-5p-modified adipose mesenchymal stem cells improve wound healing by targeting MAP2K4

doi: 10.3389/fbioe.2025.1662095

Figure Lengend Snippet: The protein expression of MAP2K4, COL1A1, α-SMA, and TNF-α in the different groups, determined by Western blot. N = 3. *: P < 0.05, vs. control; # : P < 0.05, vs. model; $ : vs. AMSCs-agomir-Exo.

Article Snippet: After blocking with 5% skim milk at 37 °C for 2 h, the membranes were incubated the primary antibodies, including anti-CD63 antibody (1: 1,000, Abclonal, Wuhan, China), anti-CD81 antibody (1: 1,000, Abclonal), anti-HSP70 antibody (1: 1,000, Proteintech, Wuhan, China), anti-calnexin antibody (1: 1,000, Proteintech), anti-MAP2K4 antibody (1: 1,000, Proteintech), anti-COL1A1 antibody (1: 1,000, NOVUS, St. Louis, MO, United States), anti-α-SMA antibody (1: 1,000, Cell Signaling Technology, Boston, MA, United States), anti-TNF-α antibody (1: 1,000, Cell Signaling Technology), and anti-GAPDH antibody (1: 10,000, Proteintech).

Techniques: Expressing, Western Blot, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Exosomes derived from miR-26a-5p-modified adipose mesenchymal stem cells improve wound healing by targeting MAP2K4

doi: 10.3389/fbioe.2025.1662095

Figure Lengend Snippet: Roles and potential mechanisms of MAP2K4 in wound healing in mice. (A) The wound healing process in the skin defect mice treated with si-NC and si-MAP2K4 at days 0, 4, 8, and 12. N = 6. (B) Quantification analysis of wound healing rate in the skin defect mice treated with si-NC and si-MAP2K4 at days 4, 8, and 12. N = 6. *: P < 0.05, vs. si-NC. (C) The mRNA expression of Map2k4 , Col1a1 , α-Sma , and Tnf-α in the skin defect mice treated with si-NC and si-MAP2K4, measured by RT-qPCR. N = 3. *: P < 0.05, vs. si-NC. (D) The level of miR-26a-5p in the skin tissues of different mice. N = 3. *: P < 0.05, vs. si-NC. (E) The protein expression of MAP2K4, COL1A1, α-SMA, and TNF-α in the skin defect mice treated with si-NC and si-MAP2K4, detected by Western blot. N = 3. *: P < 0.05, vs. si-NC.

Article Snippet: After blocking with 5% skim milk at 37 °C for 2 h, the membranes were incubated the primary antibodies, including anti-CD63 antibody (1: 1,000, Abclonal, Wuhan, China), anti-CD81 antibody (1: 1,000, Abclonal), anti-HSP70 antibody (1: 1,000, Proteintech, Wuhan, China), anti-calnexin antibody (1: 1,000, Proteintech), anti-MAP2K4 antibody (1: 1,000, Proteintech), anti-COL1A1 antibody (1: 1,000, NOVUS, St. Louis, MO, United States), anti-α-SMA antibody (1: 1,000, Cell Signaling Technology, Boston, MA, United States), anti-TNF-α antibody (1: 1,000, Cell Signaling Technology), and anti-GAPDH antibody (1: 10,000, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

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: Ecotoxicology and environmental safety

Article Title: Hexavalent chromium triggers hepatocytes premature senescence via the GATA4/NF-κB signaling pathway mediated by the DNA damage response.

doi: 10.1016/j.ecoenv.2022.113645

Figure Lengend Snippet: Fig. 3. GATA4 regulated NF-κB activation in Cr(VI)-triggered premature senescence. (A) The expressions of GATA4/NF-κB-related proteins including GATA4, TRAF3IP2, p-p65, p65, p-IκBα and IκBα were detected by western blotting. (B) western blotting and qRT-PCR were used to confirm GATA4 knockdown in L02 hepatocytes. (C) SA-β-gal staining and western blotting were used to determine the degree of cellular senescence in each group. (D) The protein levels of GATA4/NF- κB- associated proteins including GATA4, TRAF3IP2, p-p65, p65, p-IκBα and IκBα. * p < 0.05, * * p < 0.01, * ** p < 0.001 compared with the relative control.

Article Snippet: Primary antibodies of Fibronectin 1 (FN1) (15613–1-AP), GATA4 (19530–1-AP), TRAF3 Interacting Protein 2 (TRAF3IP2) (26692–1-AP), Y. Ma et al. Ecotoxicology and Environmental Safety 239 (2022) 113645 Y. Ma et al. Ecotoxicology and Environmental Safety 239 (2022) 113645 NF-κB (p65) (10745–1-AP), and senescence marker protein-30 (SMP30) (17947–1-AP) were purchased from proteintech (Wuhan, China); Phospho-NF-κB p65 (p-p65) (Ser536) (93H1), IκBα (44D4), PhosphoIκBα (p-IκBα) (Ser32) (14D4), Atg5 (D5G3) and Atg7 (D12B11) were obtained from Cell Signaling Technology (MA, USA).

Techniques: Activation Assay, Western Blot, Quantitative RT-PCR, Knockdown, Staining, Control

Journal: Ecotoxicology and environmental safety

Article Title: Hexavalent chromium triggers hepatocytes premature senescence via the GATA4/NF-κB signaling pathway mediated by the DNA damage response.

doi: 10.1016/j.ecoenv.2022.113645

Figure Lengend Snippet: Fig. 5. Cr(VI) triggered premature senescence via GATA4/NF-κB signaling pathway in vivo. (A) The degree of premature senescence in the liver tissue of mice in each group were detected by using SA-β-Gal staining kit. (B) Levels of senescence-related proteins as well as GATA4 and TRAF3IP2 were detected by western blotting. (C) The degree of senescence in the liver tissue of mice in each group after the exposure of Cr(VI) with or without PDTC were measured by SA-β-Gal staining. (D-E) Levels of senescence-related proteins and GATA4/NF-κB-related proteins expression. * p < 0.05, * * p < 0.01, * ** p < 0.001 compared with the control group.

Article Snippet: Primary antibodies of Fibronectin 1 (FN1) (15613–1-AP), GATA4 (19530–1-AP), TRAF3 Interacting Protein 2 (TRAF3IP2) (26692–1-AP), Y. Ma et al. Ecotoxicology and Environmental Safety 239 (2022) 113645 Y. Ma et al. Ecotoxicology and Environmental Safety 239 (2022) 113645 NF-κB (p65) (10745–1-AP), and senescence marker protein-30 (SMP30) (17947–1-AP) were purchased from proteintech (Wuhan, China); Phospho-NF-κB p65 (p-p65) (Ser536) (93H1), IκBα (44D4), PhosphoIκBα (p-IκBα) (Ser32) (14D4), Atg5 (D5G3) and Atg7 (D12B11) were obtained from Cell Signaling Technology (MA, USA).

Techniques: In Vivo, Staining, Western Blot, Expressing, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Fluorofenidone protects against acute liver failure in mice by regulating MKK4/JNK pathway.

doi: 10.1016/j.biopha.2023.114844

Figure Lengend Snippet: Fig. 5. AKF-PD inhibited the early activation of MKK4/JNK. (A) Representative Western blot images of phospho-MKK4 and phospho-JNK in liver tissue. (B) Semi- quantitative analysis of phospho-MKK4 (n = 4/group). (C) Semi-quantitative analysis of phospho-JNK (n = 4/group).

Article Snippet: The antibody against GAPDH (#60004–1), MKK4 (17340–1-AP), phospho-ASK1 (28201–1-AP), ASK1 (28846–1-AP) and SH3 homology-associated BTK-binding protein (SAB, 11127–2-AP) were bought from Proteintech (Wuhan, China).

Techniques: Activation Assay, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Fluorofenidone protects against acute liver failure in mice by regulating MKK4/JNK pathway.

doi: 10.1016/j.biopha.2023.114844

Figure Lengend Snippet: Fig. 6. AKF-PD protects against APAP-induced acute liver injury by regulating the MKK4/JNK pathway. Primary hepatocyte isolated from mice were pretreated with culture medium or AKF-PD (400 μg/mL) for 24 h. Following this, cells were exposed to APAP (10 mM) alone or in combination with anisomycin (10 μM) or SP600125 (10 μM) for 3 h. (A) Representative Western blot images of phospho-MKK4 and phospho-JNK in primary hepatocytes. (B) Semi-quantitative analysis of phospho-MKK4 (n = 3/group). (C) Semi-quantitative analysis of phospho-JNK (n = 3/group). Ten-week-old male C57BL/6 J mice were intraperitoneally injected with anisomycin (20 mg/kg), SP600125 (15 mg/kg) or vehicle (5%DMSO), then orally administered AKF-PD (250 mg/kg) or vehicle (0.5% CMC-Na) 30 min prior to intraperitoneal injection of acetaminophen (APAP; 400 mg/kg) (n = 5/group). The liver tissue and plasma were collected 12 h after APAP challenge. (D) Repre sentative images of H&E staining in liver tissue (20 × magnification, scale bar = 500 µm and 100 × magnification, scale bar = 100 µm). (E) The liver necrosis scores of HE staining. An overall score for each slice was computed based on 5 randomly selected scopes under 100 × magnification. (F-G) ALT and AST levels in serum. (H) Representative Western blot images of phospho-MKK4 and phospho-JNK in primary hepatocytes. (I) Semi-quantitative analysis of phospho-MKK4 (n = 4/group). (K) Semi-quantitative analysis of phospho-JNK (n = 4/group). Data are represented as mean ± SEM. *p < 0.05, * *p < 0.01, and * **p < 0.001.

Article Snippet: The antibody against GAPDH (#60004–1), MKK4 (17340–1-AP), phospho-ASK1 (28201–1-AP), ASK1 (28846–1-AP) and SH3 homology-associated BTK-binding protein (SAB, 11127–2-AP) were bought from Proteintech (Wuhan, China).

Techniques: Isolation, Western Blot, Injection, Clinical Proteomics, Staining

Journal: Frontiers in Physiology

Article Title: RARRES1 attenuates H 2 O 2 -induced RPE cell injury and inhibits choroidal neovascularization

doi: 10.3389/fphys.2025.1641653

Figure Lengend Snippet: Mechanistic Role of RARRES1 in ARPE-19 Cells Under Oxidative Stress. (A) Western blot analysis revealed that knockdown of RARRES1 in ARPE-19 cells under oxidative stress conditions led to activation of phosphorylated JNK (p-JNK) and suppression of Nrf2 and SIRT1 expression. (B) In contrast, RARRES1 overexpression reversed these effects, suggesting a protective regulatory mechanism. Protein levels were normalized to β-actin, and mRNA levels to GAPDH. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ns = not significant.

Article Snippet: The primary antibodies include RARRES1 (sc-390461, Santa Cruz), phospho-JNK (HY- P80831 , MCE), JNK (HY- P80197 , MCE), phospho-ERK (HY- P81156 , MCE), ERK (HY- P80393 , MCE), Nrf-2 (80593-1-RR, Proteintech), SIRT-1 (HY- P80319 , MCE), Occludin (502601, ZEN), β-actin (HRP-66009, Proteintech).

Techniques: Western Blot, Knockdown, Activation Assay, Expressing, Over Expression