Journal: World Journal of Stem Cells

Article Title: Effects of interleukin-10 treated macrophages on bone marrow mesenchymal stem cells via signal transducer and activator of transcription 3 pathway

doi: 10.4252/wjsc.v16.i5.560

Figure Lengend Snippet: The cytotoxicity of the special inhibitor Tyrphostin AG490. Analysis of the macrophage activity showed no significantly influences in the group with phosphorylated signal transducer and activator of transcription 3 inhibitor Tyrphostin AG490 compared with the rest of the groups by using Cell Counting Kit-8. IL-10: Interleukin-10.

Article Snippet: They were then treated differently and grouped as follows: IL-10 (100 ng/mL) was added to macrophages in the experimental group, while Tyrphostin AG490 (10 μM), a specific inhibitor of STAT3 phosphorylation (MCE, United States), was added to macrophages in another group in addition to IL-10.

Techniques: Activity Assay, Cell Counting

Journal: World Journal of Stem Cells

Article Title: Effects of interleukin-10 treated macrophages on bone marrow mesenchymal stem cells via signal transducer and activator of transcription 3 pathway

doi: 10.4252/wjsc.v16.i5.560

Figure Lengend Snippet: The effect of interleukin-10 treatment on the activation of signal transducer and activator of transcription 3 in macrophages. A: Western analysis of signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 protein in macrophages treated with/without interleukin-10. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a loading control. Tyrphostin AG490 was used at 10 μM concentration in culture medium as a phosphorylation specific inhibitor of STAT3. Data are representative of three independent experiments; B: Density values were measured by using Image J software for the representative blot shown. n = 3 biological replicates. Data are represented as mean ± SD, statistically significant difference at the levels as a P < 0.05 and b P < 0.01. IL-10: Interleukin-10; p-STAT3: Phosphorylated signal transducer and activator of transcription 3.

Article Snippet: They were then treated differently and grouped as follows: IL-10 (100 ng/mL) was added to macrophages in the experimental group, while Tyrphostin AG490 (10 μM), a specific inhibitor of STAT3 phosphorylation (MCE, United States), was added to macrophages in another group in addition to IL-10.

Techniques: Activation Assay, Western Blot, Control, Concentration Assay, Phospho-proteomics, Software

Journal: World Journal of Stem Cells

Article Title: Effects of interleukin-10 treated macrophages on bone marrow mesenchymal stem cells via signal transducer and activator of transcription 3 pathway

doi: 10.4252/wjsc.v16.i5.560

Figure Lengend Snippet: The effect of interleukin-10 acting on macrophages after the inhibition of signal transducer and activator of transcription 3. A: Cell Counting Kit-8 assay of the bone marrow mesenchymal stem cells (BMSCs) proliferation after co-cultured with different groups of macrophages: M0, M0 + interleukin-10 (IL-10), M0 + IL-10 + Tyrphostin AG490 for 1, 3, 5, and 7 d; B: Real-time polymerase chain reaction (PCR) analysis for Alkaline phosphatase mRNA level in different groups of BMSCs (groups as above); C: Real-time PCR analysis for osteocalcin mRNA level in different groups of BMSCs (groups as above); D: Real-time PCR analysis for recombinant runt related transcription factor 2 mRNA level in different groups of BMSCs (groups as above). The expression levels were normalized to the expression of glyceraldehyde-3-phosphate dehydrogenase; E: Quantitative measurement of calcium mineral deposition was stained by Alizarin Red S staining after 14 d of osteogenic induction on BMSCs. n = 3 biological replicates. Data are represented as mean ± SD, statistically significant difference at the levels as a P < 0.05 and b P < 0.01. Alp : Alkaline phosphatase; Ocn : Osteocalcin; Runx2 : Recombinant runt related transcription factor 2; Gapdh : Glyceraldehyde-3-phosphate dehydrogenase; IL-10: Interleukin-10.

Article Snippet: They were then treated differently and grouped as follows: IL-10 (100 ng/mL) was added to macrophages in the experimental group, while Tyrphostin AG490 (10 μM), a specific inhibitor of STAT3 phosphorylation (MCE, United States), was added to macrophages in another group in addition to IL-10.

Techniques: Inhibition, Cell Counting, Cell Culture, Real-time Polymerase Chain Reaction, Recombinant, Expressing, Staining

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