Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: Network pharmacology and molecular docking of the main components of JQP with the STAT3 protein molecule. A. Venn diagram of candidate target genes of JQP and AS. B. Protein-protein interaction network, the darker the color, the higher the importance. C. Degree screening of the top 20 gene targets. D. Cytoscape 3.7.1 The software draws the active ingredient-target gene network of TCM, in which the yellow square represents the target gene, the green circle represents the traditional Chinese medicine, and the green square represents the active ingredient of the traditional Chinese medicine. E. Bubble chart of KEGG pathway enrichment analysis of candidate AS targets. The size of the dots indicates the number of selected genes, and the color indicates the p value of the enrichment analysis. F. Visualization of molecular docking of active ingredients and core protein targets in the core formulation. (a) Docking diagram of quercetin (MOL000098) and STAT3. (b) Docking diagram of kaempferol (MOL000422) and STAT3. (c) Docking diagram of wogonin (MOL000173) and STAT3. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Software, Formulation

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: Effects of NONHSAT227927.1 on cell viability, inflammatory cytokines and inflammatory pathways.A, Screening of NONHSAT227927.1 small interfering RNA model. B, CCK-8 method was used to detect the cell viability of AS-FLS. C, RT-qPCR was used to detect the expression of NONHSAT227927.1 in AS-FLS. D-G, ELISA was used to detect the levels of IL-6, IL-17, TNF-α, and IL-10 in AS-FLS. H–I, RT-qPCR was used to detect the levels of JAK2 and STAT3 in AS-FLS. J, WB was used to detect Levels of JAK2, STAT, p -JAK2 and p-STAT3 in AS-FLS. K, IF was used to detect the expression of JAK2 and STAT3 in AS-FLS. All experiments were repeated three times.a: ov-NC, b: ov-NONHSAT227927.1, c: si-NC, d: si-NONHSAT227927.1. ***P < 0.001.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Small Interfering RNA, CCK-8 Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: NONHSAT227927.1/JAK2/STAT3 combination regulates the activity of AS-FLS and the release of inflammatory factors. A, CCK-8 method was used to detect the cell viability of AS-FLSs. B-E, ELISA method was used to detect the levels of IL-6, IL-17, TNF-α, and IL-10 in AS-FLS. FG, RT-qPCR was used to detect Levels of JAK2 and STAT3 in AS-FLS. H, WB was used to detect the levels of JAK2, STAT, p -JAK2 and p-STAT3 in AS-FLS. I, IF was used to detect the expression of JAK2 and STAT3 in AS-FLS. All experiments were repeated three times. a: FLS, b: AG490, c: ov-NC, d: ov-NONHSAT227927.1, e: AG490 + ov-NONHSAT227927.1.*P < 0.05,**P < 0.01,***P < 0.001.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Activity Assay, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: JQP reversed the effects of NONHSAT227927.1 overexpression on AS-FLS viability, inflammatory factors and JAK2/STAT3 pathway. A, CCK-8 method was used to screen the optimal concentration of JQP-containing serum. B. Use CCK-8 method to detect the cell viability of AS-FLSs. C. RT-qPCR detects the expression of NONHSAT227927.1. D-G used ELISA to detect the levels of IL-6, IL-17, TNF-α, and IL-10. HI, RT-qPCR was used to detect the expression of JAK2 and STAT3. J. WB detects the expression of JAK2, STAT3, p -JAK2 and p-STAT3 proteins. K. Immunofluorescence method detects the expression of p -JAK2 and p-STAT3 proteins. a: FLS, b: NS, c: JQP, d: ov-NC, e: ov-NONHSAT227927.1, f: ov-NONHSAT227927.1 + JQP. The data is displayed in the form of the average value plus or minus the standard deviation. The experiments were conducted thrice.*P < 0.05,**P < 0.01,***P < 0.001.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques: Over Expression, CCK-8 Assay, Concentration Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Standard Deviation

Journal: Heliyon

Article Title: Jianpi Qingre Tongluo Decoction exerted an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis

doi: 10.1016/j.heliyon.2024.e34634

Figure Lengend Snippet: JQP exerts an anti-inflammatory effect on AS by inhibiting the NONHSAT227927.1/JAK2/STAT3 axis.

Article Snippet: The JAK2/STAT3 pathway inhibitor AG-490 (10 μm) (MCE, No.: HY-12000) was applied to AS-FLSs for a duration of 24 h. To overexpress NONHSAT227927.1, the NONHSAT227927.1 gene was introduced into the pcDNA3.1 plasmid (GenePharma, Shanghai, China).

Techniques:

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation exacerbated mortality in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, 50, or 100 µM AG490, followed by co-treatment with 0 or 10 µM MeHg. Cytotoxicity (LDH release) was measured using LDH at 3 h ( A ), 6 h ( B ), and 24 h ( C ) of MeHg addition. Cell viability was assessed using MTT at 3 h ( D ), 6 h ( E ), and 24 h ( F ) of MeHg addition. Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation increases oxidative stress in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, 50, or 100 µM AG490 ( A - E ) or with 0, 3, or 30 µM C188-9 ( F - J ), followed by co-treatment with 0 or 10 µM MeHg. ROS production was measured using CM-H2DCFDA probe after MeHg addition at 30 min ( A , F ), 1 h ( B , G ), 3 h ( C , H ), 6 h ( D , I ), and 24 h ( E , J ). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation increases mitochondrial oxidative stress in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, 50, or 100 µM AG490 ( A - D ) or with 0, 3, or 30 µM C188-9 ( F - H ), followed by co-treatment with 0 or 10 µM MeHg. Mitochondrial superoxide production was assessed using mitoSOX fluorescent staining, after MeHg addition at 30 min ( A , E ), 1 h ( B , F ), 3 h ( C , G ), and 6 h ( D , H ). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Staining, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation exacerbates GSH depletion in C8-D1A astrocytic cells. Cells were pretreated with 0 or 100 µM AG490 ( A , B , C ) or with 0 or 30 µM C188-9 ( D , E , F ), followed by co-treatment with 0 or 10 µM MeHg. Total GSH ( A , D ) reduced GSH ( B , C ), and GSH/GSSG ratio were measured at 24 h after adding MeHg. Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: STAT3 inhibition induces aberrant antioxidant enzyme expression in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, or 100 µM AG490 or with 0 or 30 µM C188-9, followed by 24 h of co-treatment with 0 or 10 µM MeHg. Protein levels of phosphorylated STAT3 to total STAT3 ratio (pSTAT3/STAT3) ( A , E ), HO-1 ( B , F ), and Nrf2 ( C , G ) were measured by western blot. Representative densitometry images are shown in ( D , H ). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. P < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Expressing, Western Blot, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: STAT3 inhibition induces aberrant antioxidant gene expression in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, or 100 µM AG490 or with 0 or 30 µM C188-9, followed by 24 h of co-treatment with 0 or 10 µM MeHg. Gene expression of Stat3 ( A , C ) and Socs3 ( B , D ) at 3 h, and Stat3 ( E , I ), Socs3 ( F , J ), Hmox1 ( G , K ), and Slc7a11 ( H , L ) at 24 h was measured by qPCR. Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Gene Expression, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: STAT3 inhibition did not prevent MeHg IL-6 release in C8-D1A astrocytic cells. Cells were pretreated with 0, 10 or 100 µM AG490 ( A , B ) or with 0, 3, or 30 µM C188-9 ( C , D ), followed by co-treatment with 0 or 10 µM MeHg. Il- 6 mRNA expression ( A , C ) was measured by qPCR at 3 h after adding MeHg. IL-6 release into the medium ( B , D ) was measured at 16 h after adding MeHg Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Expressing, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: SOCS1 Inhibits IL-6-Induced CD155 Overexpression in Lung Adenocarcinoma

doi: 10.3390/ijms252212141

Figure Lengend Snippet: Effect of IL-6 stimulation on CD155 expression on lung adenocarcinoma cell lines. ( A ) IL-6 stimulation induced CD155 mRNA expression in the tested cell lines. ( B ) Upon IL-6 stimulation, CD155 protein expression increased in A-549 and NCI-H1437 cells, but no change was detected in the advanced stages 3B1A and NCI-H1573 cell lines. ( C ) Results show that CD155 concentration was augmented only in the supernatant of the IL-6-stimulated A-549 cell line. ( D ) Addition of the JAK2 inhibitor AG490 prevents IL-6-induced CD155 overexpression in the A-549 and NCI-H1437 cell lines. ( A , C ) are shown as mean + SD. * p < 0.05, ** p < 0.01.

Article Snippet: In addition, to confirm that the observed effect was IL-6 dependent, the JAK2 inhibitor, Tyrphostin AG490 (Cat. No. S1143; Selleckchem, Houston, TX, USA), was added at a concentration of 15 μM prior to IL-6 addition.

Techniques: Expressing, Concentration Assay, Over Expression