sil 6r (R&D Systems)
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sil 6r
Sil 6r, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 1392 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sil 6r/product/R&D Systems
Average 97 stars, based on 1392 article reviews
Sil 6r, supplied by R&D Systems, used in various techniques. Bioz Stars score: 97/100, based on 1392 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/sil 6r/product/R&D Systems
Average 97 stars, based on 1392 article reviews
sil 6r - by Bioz Stars,
2026-05
97/100 stars
Images
1) Product Images from "Synovial fibroblast niche shapes the efficacy–safety dynamics of JAK inhibition in rheumatoid arthritis"
Article Title: Synovial fibroblast niche shapes the efficacy–safety dynamics of JAK inhibition in rheumatoid arthritis
Journal: bioRxiv
doi: 10.64898/2026.03.23.713616
Figure Legend Snippet: a-f ) In vitro experiments. We stimulated cultured rheumatoid arthritis (RA) synovial fibroblasts (SF) with 50ng/ml interleukin 6 (IL-6) or 1ng/ml tumor necrosis factor (TNF) with or without 50ng/ml soluble IL-6 receptor (sIL-6R). a, b) Basal and cytokine-induced phosphorylation of JAK1 and STAT1/3 proteins in SF stimulated with a) IL-6 + sIL-6R for 10min, n=15-16 biological replicates or b) IL-6 + sIL-6R, TNF + sIL-6R or TNF for 5h, n=13-16 biological replicates. We quantified the data as a ratio of phosphorylated versus total protein (pProtein/Protein), using optical density values of chemiluminescently-detected immunoblotted proteins. pProtein/Protein values were log10 transformed. Representative immunoblot images are shown in , and original images in Suppl. Figs. 3, 4 . c) Comparison of early (10min) versus late (5h) induction of pJAK1 and pSTAT1/3 in SF, calculated as x-fold = pProtein/Protein (stimulated) / pProtein/Protein (unstimulated) per time point; x-fold values were log10 transformed. a-c) We present experimental data using box and whisker plots with median and min to max. We compared paired experimental groups using paired t test or Wilcoxon matched-pairs signed rank test. *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001. d-f) Expression of JAK-STAT pathway-associated genes, probed on TaqMan ArrayCards, in unstimulated and stimulated (IL-6/TNF + sIL-6R) RA SF; n=11 biological replicates per experimental condition. We quantified gene expression as ΔCt; 72 genes were reliably detected across n = 33 experimental conditions. Not detected genes were excluded from analysis. d) Principal component (PC) analysis of ΔCt values using ClustVis. PC2 and PC3 separated samples according to stimulus and donor, respectively. PC1 was dominated by a single outlier sample ( Suppl. Fig. 2b). e) Similarity matrix, computed for columns (samples) using Pearson correlation in Morpheus. f) Heatmap generated in Morpheus using ΔCt gene expression values, with hierarchical clustering of genes and experimental conditions. Rows and columns were clustered using 1 − Pearson correlation with average linkage.
Techniques Used: In Vitro, Cell Culture, Phospho-proteomics, Transformation Assay, Western Blot, Comparison, Whisker Assay, Expressing, Gene Expression, Generated
![a-h ) In vitro experiments. We stimulated cultured rheumatoid arthritis synovial fibroblasts (SF) with 50ng/ml interleukin 6 (IL-6) or 1ng/ml tumor necrosis factor (TNF) in the presence or absence of 50ng/ml soluble IL-6 receptor (sIL-6R) for 10min or 5h across tofacitinib concentrations (0, 180, 1000nM). Cells were pretreated with tofacitinib for 2h. a, b) Representative immunoblot images of (un)phosphorylated JAK1 and STAT1/3 proteins as well as GAPDH (loading control) in unstimulated SF and SF, stimulated with a) IL-6 and sIL-6R for 10min. N = 9-10 biological replicates or b) IL-6 + sIL-6R or TNF ± sIL-6R for 5h, n=7-10 biological replicates. Original immunoblot images are shown in Suppl. Figs. 3, 4 . c, d ) We quantified immunoblot data as a ratio of phosphorylated versus total protein, using optical density values of chemiluminesce-detected immunoblotted proteins. Data are presented using box and whisker plots with median and min to max. To compare the effects of different experimental conditions (stimulated, unstimulated) at individual tofacitinib concentrations, we used c) paired t-test / Wilcoxon matched-pairs signed-rank test or d) RM one-way ANOVA with Šídák’s multiple-comparisons test / Friedman test with Dunn’s multiple-comparisons test. Biologically meaningful pairwise comparisons were performed. To evaluate the effects of different tofacitinib concentrations (0, 180, 1000um) within each experimental condition, we performed RM one-way ANOVA with Tukey’s multiple-comparisons test or Friedman test with Dunn’s multiple-comparisons test. e, f) Percent inhibition of pJAK1 and pSTAT1/3, normalised to total JAK1 and STAT1/3, respectively, in IL-6 + sIL-6R-treated SF at e) 10min and f) 5h time points. We compared tofacitinib concentration effects using paired t-test or Wilcoxon matched-pairs signed-rank test. g, h) Residual pJAK1 and pSTAT1/3 phosphorylation, normalised to total JAK1 and STAT1/3, respectively, in tofacitinib-treated cytokine-stimulated SF. Log10 [x-fold(s,d)] = log10 [pProtein/Protein(s,d) / pProtein/Protein ( 0,0 ) ], where s denotes stimulus (IL-6 + sIL-6R, TNF + sIL-6R, TNF), d represents tofacitinib concentration (0, 180, 1000nM) and baseline log10 [x-fold(0,0] equals 0 in unstimulated SF at 0 uM tofacitinib. Data are shown using box and whisker plots with median and min to max. To compare tofacitinib concentration effects within individual stimuli, we performed RM one-way ANOVA with Tukey’s multiple-comparisons test or the Friedman test with Dunn’s multiple-comparisons test. To compare log10 [x-fold(s,d)] values to baseline log10[x-fold] we used a one-sample t-test or Wilcoxon test. p-value was adjusted for multiple comparisons using the Bonferroni test. *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001.](https://bio-rxiv-images-cdn.bioz.com/dois_ending_with_16/10__64898_slash_2026__03__23__713616/10__64898_slash_2026__03__23__713616___F7.large.jpg "... presence or absence of 50ng/ml soluble IL-6 receptor (sIL-6R) for 10min or 5h across tofacitinib concentrations (0, ...")
Figure Legend Snippet: a-h ) In vitro experiments. We stimulated cultured rheumatoid arthritis synovial fibroblasts (SF) with 50ng/ml interleukin 6 (IL-6) or 1ng/ml tumor necrosis factor (TNF) in the presence or absence of 50ng/ml soluble IL-6 receptor (sIL-6R) for 10min or 5h across tofacitinib concentrations (0, 180, 1000nM). Cells were pretreated with tofacitinib for 2h. a, b) Representative immunoblot images of (un)phosphorylated JAK1 and STAT1/3 proteins as well as GAPDH (loading control) in unstimulated SF and SF, stimulated with a) IL-6 and sIL-6R for 10min. N = 9-10 biological replicates or b) IL-6 + sIL-6R or TNF ± sIL-6R for 5h, n=7-10 biological replicates. Original immunoblot images are shown in Suppl. Figs. 3, 4 . c, d ) We quantified immunoblot data as a ratio of phosphorylated versus total protein, using optical density values of chemiluminesce-detected immunoblotted proteins. Data are presented using box and whisker plots with median and min to max. To compare the effects of different experimental conditions (stimulated, unstimulated) at individual tofacitinib concentrations, we used c) paired t-test / Wilcoxon matched-pairs signed-rank test or d) RM one-way ANOVA with Šídák’s multiple-comparisons test / Friedman test with Dunn’s multiple-comparisons test. Biologically meaningful pairwise comparisons were performed. To evaluate the effects of different tofacitinib concentrations (0, 180, 1000um) within each experimental condition, we performed RM one-way ANOVA with Tukey’s multiple-comparisons test or Friedman test with Dunn’s multiple-comparisons test. e, f) Percent inhibition of pJAK1 and pSTAT1/3, normalised to total JAK1 and STAT1/3, respectively, in IL-6 + sIL-6R-treated SF at e) 10min and f) 5h time points. We compared tofacitinib concentration effects using paired t-test or Wilcoxon matched-pairs signed-rank test. g, h) Residual pJAK1 and pSTAT1/3 phosphorylation, normalised to total JAK1 and STAT1/3, respectively, in tofacitinib-treated cytokine-stimulated SF. Log10 [x-fold(s,d)] = log10 [pProtein/Protein(s,d) / pProtein/Protein ( 0,0 ) ], where s denotes stimulus (IL-6 + sIL-6R, TNF + sIL-6R, TNF), d represents tofacitinib concentration (0, 180, 1000nM) and baseline log10 [x-fold(0,0] equals 0 in unstimulated SF at 0 uM tofacitinib. Data are shown using box and whisker plots with median and min to max. To compare tofacitinib concentration effects within individual stimuli, we performed RM one-way ANOVA with Tukey’s multiple-comparisons test or the Friedman test with Dunn’s multiple-comparisons test. To compare log10 [x-fold(s,d)] values to baseline log10[x-fold] we used a one-sample t-test or Wilcoxon test. p-value was adjusted for multiple comparisons using the Bonferroni test. *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001.
Techniques Used: In Vitro, Cell Culture, Western Blot, Control, Whisker Assay, Inhibition, Concentration Assay, Phospho-proteomics
Figure Legend Snippet: a, b ) In vitro experiments. We stimulated cultured rheumatoid arthritis (RA) synovial fibroblasts (SF) with 50ng/ml interleukin 6 (IL-6) or 1ng/ml tumor necrosis factor (TNF) ± 50ng/ml soluble IL-6 receptor (sIL-6R) for 24h. N = 6-11 biological replicates per experimental condition. Gene expression was quantified using qPCR and calculated as ΔCt. We show ΔCt using box and whisker plots with median and min to max. To evaluate differences among preselected biologically meaningful pairs of experimental conditions (unstimulated, stimulated), we used RM one-way ANOVA with Šídák’s multiple comparisons test or Friedman test with Dunn’s multiple comparisons test. a) Genes upregulated in the presence of both IL-6 + sIL-6R and TNF ± sIL-6R. b) Genes selectively upregulated with TNF ± sIL-6R. c, d) In vitro experiments. We stimulated cultured RA SF with 50ng/ml IL-6, 0.1 or 1ng/ml TNF in the presence or absence of 50ng/ml sIL-6R. Secreted c) IL-6 (n=12-14 biological replicates) and d) IL-8 (n=5-6 biological replicates) proteins were analysed 24h after stimulation using ELISA. Protein concentration values (pg/ml) were log10-normalised. Data are presented using box and whiskers plots with median and min to max. To evaluate differences among preselected biologically meaningful pairs of experimental conditions (unstimulated, stimulated), we used mixed-effects analysis with Šídák’s multiple comparisons test. *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001.
Techniques Used: In Vitro, Cell Culture, Gene Expression, Whisker Assay, Enzyme-linked Immunosorbent Assay, Protein Concentration
Figure Legend Snippet: a) In vitro experiments. We stimulated cultured rheumatoid arthritis (RA) synovial fibroblasts (SF) with 50ng/ml interleukin 6 (IL-6) or 1ng/ml tumor necrosis factor (TNF 1) in the presence or absence of 50ng/ml soluble IL-6 receptor (sIL-6R) for 24h across tofacitinib concentrations (0, 180, 1000nM). Cells were pretreated with tofacitinib for 2h. Gene expression was quantified using qPCR and calculated as ΔCt. N = 5-10 biological replicates per experimental condition. Data are presented using box and whiskers plots with median and min to max. To compare unstimulated and stimulated experimental conditions in the absence of tofacitinib, we performed RM one-way ANOVA with Šídák’s multiple-comparisons test or the Friedman test with Dunn’s multiple-comparisons test. Biologically meaningful pairwise comparisons were performed. To compare tofacitinib concentration effects within individual experimental conditions (unstimulated and stimulated), we used RM one-way ANOVA with Tukey’s multiple-comparisons test or the Friedman test with Dunn’s multiple-comparisons test. b, c) In vitro experiments. We stimulated cultured RA SF with 50ng/ml IL-6, 0.1 or 1ng/ml TNF in the presence or absence of 50ng/ml sIL-6R across tofacitinib concentrations (0, 80, 180, 1000nM). Cells were pretreated with tofacitinib for 2h. Secreted b) IL-6 (n=11-14 biological replicates) and c) IL-8 (n=5-6 biological replicates) proteins were analysed 24h after stimulation using ELISA. Data are presented using box and whiskers plots with median and min to max. x-fold = Interleukin(s,d) / Interleukin(s,0) , where Interleukin denotes the cytokine (IL-6, IL-8), s denotes the experimental condition (unstimulated or stimulated), d denotes the tofacitinib concentration (0, 80, 180, 1000nM), and x-fold = 1 represents the baseline x-fold, calculated for the 0nM tofacitinib for each experimental condition. To compare x-fold values to baseline x-fold = 1, we performed a one-sample t-test or Wilcoxon test. p-values were adjusted for multiple comparisons with the Bonferroni test. To compare x-fold values across tofacitinib concentrations within individual experimental conditions, we used RM one-way ANOVA with Tukey’s multiple comparisons test or Friedman test with Dunn’s multiple comparisons test. *p<0.05, **p<0.01, *** p<0.001, ****p<0.0001.
Techniques Used: In Vitro, Cell Culture, Gene Expression, Concentration Assay, Enzyme-linked Immunosorbent Assay
Figure Legend Snippet: a) Schematic of the tofacitinib washout experiment in cultured rheumatoid arthritis synovial fibroblasts (SF); created with BioRender. We pretreated SF with tofacitinib (180, 1000nM) or equal volumes of DMSO for 2h. Thereafter, we stimulated SF with 50ng/ml interleukin 6 (IL-6) and 50ng/ml soluble IL-6 receptor (sIL-6R) in the presence of tofacitinib (180, 1000nM) or DMSO for 10min or 5h. Finally, we either directly lysed SF, washed out supernatants with/without tofacitinib re-supplementation, or continued treatment for an additional 10min. b) Representative immunoblot images of (un)phosphorylated JAK1 and STAT1/3 proteins and GAPDH (loading control) in SF are shown across experimental conditions. Original immunoblot images are depicted in Suppl. Figs. 5, 6 . We quantified experimental data from c) 10min and d) 5h washout experiments as a ratio of phosphorylated versus total protein, using optical density values of chemiluminesce-detected immunoblotted proteins. N=3 biological replicates per experimental condition.
Techniques Used: Cell Culture, Western Blot, Control