Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Vaccarin Improves Myocardial Ischemia–Reperfusion Injury by Attenuating Oxidative Stress and Ferroptosis Through Reducing NOX4 ‐Modulated JAK2 / STAT3 Pathway Activation

doi: 10.1002/kjm2.70226

Figure Lengend Snippet: Vaccarin prevents the activation of JAK2/STAT3, which is mediated by NOX4. (A–C) Inhibitory effect of vaccarin on JAK2/STAT3 signaling in I/R mice ( n = 6). Statistical analyses were performed using one‐way ANOVA, followed by Tukey's post hoc test. (D, E) NOX4‐overexpressing H9c2 cells were constructed through transfection with pcDNA/NOX4. The experiment was repeated three times. Statistical analyses were performed using one‐way ANOVA, followed by Dunnett's post hoc test. (F–H) H9c2 cells were transfected with an empty pcDNA3.1 plasmid (vector) or pcDNA/NOX4. After 24 h of transfection, the cells were pretreated with vaccarin (10 μM) and/or AG490 (10 μM) for 30 min, followed by H/R induction. Western blotting was conducted to determine the expression of p‐JAK2, JAK2, p‐STAT3, and STAT3. The experiment was repeated three times. Statistical analyses were performed using one‐way ANOVA, followed by Tukey's post hoc test. Data are presented as mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: To inhibit the JAK2/STAT3 pathway, JAK2 inhibitor AG490 (10 μM; HY‐12000, purity 99.86%, MedChemExpress) was used.

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

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Vaccarin Improves Myocardial Ischemia–Reperfusion Injury by Attenuating Oxidative Stress and Ferroptosis Through Reducing NOX4 ‐Modulated JAK2 / STAT3 Pathway Activation

doi: 10.1002/kjm2.70226

Figure Lengend Snippet: The inhibitory effects of vaccarin on H/R‐induced oxidative stress and ferroptosis in cardiomyocytes are mediated by the NOX4/JAK2/STAT3 signaling pathway. H9c2 cells were transfected with an empty pcDNA3.1 plasmid (vector) or pcDNA/NOX4. After 24 h of transfection, the cells were pretreated with vaccarin (10 μM) and/or AG490 (10 μM) for 30 min, followed by H/R induction. (A–D) ROS and MDA production and SOD activity were determined. The experiment was repeated three times in triplicate. (E–G) The levels of Fe 2+ , GSSG, and GSH were determined. The experiment was repeated three times in triplicate. (H–J) The expression levels of SLC7A11 and GPX4 were determined. The experiment was repeated three times. Statistical analyses were performed using one‐way ANOVA, followed by Tukey's post hoc test. Data are presented as mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: To inhibit the JAK2/STAT3 pathway, JAK2 inhibitor AG490 (10 μM; HY‐12000, purity 99.86%, MedChemExpress) was used.

Techniques: Transfection, Plasmid Preparation, Activity Assay, Expressing

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Vaccarin Improves Myocardial Ischemia–Reperfusion Injury by Attenuating Oxidative Stress and Ferroptosis Through Reducing NOX4 ‐Modulated JAK2 / STAT3 Pathway Activation

doi: 10.1002/kjm2.70226

Figure Lengend Snippet: Mechanisms underlying the cardioprotective role of vaccarin in MI/R injury. Vaccarin suppressed the myocardial I/R‐induced NOX4 expression, thus inactivating the JAK2/STAT3 signaling. The suppression of NOX4‐mediated JAK2/STAT3 signaling contributed to the antioxidative and antiferroptotic effects of vaccarin in MI/R injury.

Article Snippet: To inhibit the JAK2/STAT3 pathway, JAK2 inhibitor AG490 (10 μM; HY‐12000, purity 99.86%, MedChemExpress) was used.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: p-JAK2/JAK2 and p-STAT3/STAT3 were highly expressed in OLP. AG490 and RPM inhibited KCs proliferation. AG490 and RPM inhibited homing migration of OLP IELs. (A–C) p-JAK2/JAK2 and p-STAT3/STAT3 were significantly upregulated in OLP compared to controls (n = 6), as detected by western blot. (D) In the MTT assay, at 48 h, the OD450 values of the AG490 group, RPM group, and AG490+RPM group were significantly lower than those of the Control group. Compared with the AG490 group and RPM group, the OD450 values of the AG490+RPM group were significantly reduced. Additionally, the OD450 values of the AG490 group were significantly lower than those of the RPM group. (E) Compared with the control group, the AG490 group and RPM group exhibited a significant reduction in the number of migration tracks, while the AG490+RPM group showed a significantly reduced number of tracks. (F, G) Compared with the control group, the AG490 group and RPM group demonstrated a significant decrease in both speed and maximum displacement, and the AG490+RPM group exhibited a significant reduction in both speed and maximum displacement. * p < 0.05; ** p < 0.01; *** p < 0.001. JAK2, Janus kinase 2; p-JAK2, Phosphorylated Janus kinase 2; STAT3, Signal transducer and activator of transcription 3; p-STAT3, Phosphorylated signal transducer and activator of transcription 3; OD, Optical density; AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Migration, Western Blot, MTT Assay, Control

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: AG490 and RPM inhibited the expression of apoptosis-related proteins Bax and caspase-3 and promoted the expression of Bcl-2, as measured by ELISA. (a), compared with control, p < 0.05; (b), compared with AG490 group, p < 0.05; (c), compared with RPM group, p < 0.05. AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib; Bax, Bcl-2 associated X protein; Bcl-2, B-cell lymphoma/leukemia-2 gene; caspase-3, Cysteine-dependent aspartate-specific protease-3.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: AG490 and RPM synergistically inhibited the phosphorylation of JAK2/STAT3. (A, B) AG490 and RPM selectively inhibited JAK2/STAT3 phosphorylation. * p < 0.05; ** p < 0.01; *** p < 0.001. (C) JAK2 and p-JAK2 were downregulated in AG490 and AG490+RPM group. (D) STAT3 and p-STAT3 were downregulated in RPM and AG490+RPM group. (E) Compared with the control group, the protein ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly increased in the AG490 group, RPM group, and AG490+RPM group. Compared with the AG490 group and RPM group, the protein ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly decreased in the AG490+RPM group. a, compared with control, p < 0.05; b, compared with AG490 group, p < 0.05; c, compared with RPM group, p < 0.05; aa, compared with control, p < 0.01. AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib; JAK2, Janus kinase 2; p-JAK2, Phosphorylated Janus kinase 2; STAT3, Signal transducer and activator of transcription 3; p-STAT3: Phosphorylated signal transducer and activator of transcription 3; GAPDH, Glycerol-3-phosphate dehydrogenase.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Phospho-proteomics, Control

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: The expression of ZO-1 and Occludin mRNA was upregulated after using AG490 and RPM. (A, B) Compared with the control group, the expression of ZO-1 and Occludin mRNA increased in the AG490 group. The expression of ZO-1 and Occludin mRAN in the RPM group was upregulated. The expression of ZO-1 and Occludin mRNA was significantly increased in the AG490+RPM group. Compared with the RPM group, the AG490+RPM group showed increased expression of ZO-1 and Occludin mRNA. * p < 0.05; ** p < 0.01; *** p < 0.001. (C) Schematic diagram of the regulatory network mechanism involving E-cadherin/CD103-JAK2/STAT3-ZO-1/Occludin in OLP. The binding of E-cadherin/CD103 downregulated JAK2/STAT3 phosphorylation in KCs and upregulated mucosal barrier molecules ZO-1 and Occludin, which helped maintain the integrity of the mucosal barrier. CD103, Integrin alpha-E; E-cadherin, Epithelial cadherin; JAK2, Janus kinase 2; STAT3, Signal transducer and activator of transcription 3; AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib; ZO-1, Zonula occludens protein 1; KC, Keratinocytes; IEL, Intraepithelial lymphocyte.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Expressing, Control, Binding Assay, Phospho-proteomics

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: p-JAK2/JAK2 and p-STAT3/STAT3 were highly expressed in OLP. AG490 and RPM inhibited KCs proliferation. AG490 and RPM inhibited homing migration of OLP IELs. (A–C) p-JAK2/JAK2 and p-STAT3/STAT3 were significantly upregulated in OLP compared to controls (n = 6), as detected by western blot. (D) In the MTT assay, at 48 h, the OD450 values of the AG490 group, RPM group, and AG490+RPM group were significantly lower than those of the Control group. Compared with the AG490 group and RPM group, the OD450 values of the AG490+RPM group were significantly reduced. Additionally, the OD450 values of the AG490 group were significantly lower than those of the RPM group. (E) Compared with the control group, the AG490 group and RPM group exhibited a significant reduction in the number of migration tracks, while the AG490+RPM group showed a significantly reduced number of tracks. (F, G) Compared with the control group, the AG490 group and RPM group demonstrated a significant decrease in both speed and maximum displacement, and the AG490+RPM group exhibited a significant reduction in both speed and maximum displacement. * p < 0.05; ** p < 0.01; *** p < 0.001. JAK2, Janus kinase 2; p-JAK2, Phosphorylated Janus kinase 2; STAT3, Signal transducer and activator of transcription 3; p-STAT3, Phosphorylated signal transducer and activator of transcription 3; OD, Optical density; AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Migration, Western Blot, MTT Assay, Control

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: AG490 and RPM inhibited the expression of apoptosis-related proteins Bax and caspase-3 and promoted the expression of Bcl-2, as measured by ELISA. (a), compared with control, p < 0.05; (b), compared with AG490 group, p < 0.05; (c), compared with RPM group, p < 0.05. AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib; Bax, Bcl-2 associated X protein; Bcl-2, B-cell lymphoma/leukemia-2 gene; caspase-3, Cysteine-dependent aspartate-specific protease-3.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: AG490 and RPM synergistically inhibited the phosphorylation of JAK2/STAT3. (A, B) AG490 and RPM selectively inhibited JAK2/STAT3 phosphorylation. * p < 0.05; ** p < 0.01; *** p < 0.001. (C) JAK2 and p-JAK2 were downregulated in AG490 and AG490+RPM group. (D) STAT3 and p-STAT3 were downregulated in RPM and AG490+RPM group. (E) Compared with the control group, the protein ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly increased in the AG490 group, RPM group, and AG490+RPM group. Compared with the AG490 group and RPM group, the protein ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly decreased in the AG490+RPM group. a, compared with control, p < 0.05; b, compared with AG490 group, p < 0.05; c, compared with RPM group, p < 0.05; aa, compared with control, p < 0.01. AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib; JAK2, Janus kinase 2; p-JAK2, Phosphorylated Janus kinase 2; STAT3, Signal transducer and activator of transcription 3; p-STAT3: Phosphorylated signal transducer and activator of transcription 3; GAPDH, Glycerol-3-phosphate dehydrogenase.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Phospho-proteomics, Control

Journal: Frontiers in Immunology

Article Title: Keratinocytes regulate intraepithelial lymphocytes homing and mediate mucosal barrier integrity via JAK2/STAT3 signaling in oral lichen planus

doi: 10.3389/fimmu.2026.1794867

Figure Lengend Snippet: The expression of ZO-1 and Occludin mRNA was upregulated after using AG490 and RPM. (A, B) Compared with the control group, the expression of ZO-1 and Occludin mRNA increased in the AG490 group. The expression of ZO-1 and Occludin mRAN in the RPM group was upregulated. The expression of ZO-1 and Occludin mRNA was significantly increased in the AG490+RPM group. Compared with the RPM group, the AG490+RPM group showed increased expression of ZO-1 and Occludin mRNA. * p < 0.05; ** p < 0.01; *** p < 0.001. (C) Schematic diagram of the regulatory network mechanism involving E-cadherin/CD103-JAK2/STAT3-ZO-1/Occludin in OLP. The binding of E-cadherin/CD103 downregulated JAK2/STAT3 phosphorylation in KCs and upregulated mucosal barrier molecules ZO-1 and Occludin, which helped maintain the integrity of the mucosal barrier. CD103, Integrin alpha-E; E-cadherin, Epithelial cadherin; JAK2, Janus kinase 2; STAT3, Signal transducer and activator of transcription 3; AG490, (E)-N-Benzyl-2-cyano-3-(3,4-dihydroxyphenyl)prop-2-enamide; RPM, Ruxolitinib; ZO-1, Zonula occludens protein 1; KC, Keratinocytes; IEL, Intraepithelial lymphocyte.

Article Snippet: The models were divided into four groups based on the addition of inhibitors: (1) Control group (treated with K-SFM medium containing an equal concentration of PBS); (2) AG490 group (treated with K-SFM medium containing 50 μmol/L AG490) (AG490, Cat # HY-12003, MCE); (3) RPM group (treated with K-SFM medium containing 20 nmol/L RPM) (RPM, Cat # HY-10219, MCE); and (4) AG490+RPM group (treated with K-SFM medium containing 50 μmol/L AG490 and 20 nmol/L RPM).

Techniques: Expressing, Control, Binding Assay, Phospho-proteomics