Fig. 1 . (A) RT-PCR expression analysis of IFNG , TNFA , Saa1 and Cxcl13 depicted as Cumming plots. CT values over 40 cycles and undetermined values were considered as not expressed and set to zero. (B) Plasma expression levels of human IFNγ, IL-17A, IL12p70 and TNFα detected by Luminex assay and displayed as Cumming plots. The upper part of the plot presents each data point in a swarmplot. The mean and s.d. of each group is plotted as a gapped line, where the vertical lines correspond to the mean±s.d. and the mean itself is depicted as a gap in the line (*** P ≤0.001, ** P ≤0.01, * P ≤0.05). Statistical analysis was performed with R (Kruskal–Wallis test). mRNA levels are depicted as lg(−deltaCT). RA, N =3, n =17; RA+, N =4, n =27; nonRA, N =1, n =6; nonRA+, N =1, n =6. Variations in sample sizes can be attributed to challenges encountered during RNA isolation and the exclusion of outliers during data processing. Values deviating more than sixfold from the mean were considered outliers. As indicated in the key to the right of the lower plot in B, plasma from the RA5 samples was not included in the human TNFα analysis, resulting in a decreased sample size for that plot. N =number of donors, n =total number of mice. Mice were reconstituted with PBMCs from patients with RA (RA3, n =6; RA4, n =6; RA5, n =11; RA7, n =6; RA8, n =15) and an unaffected individual (RA6, n =12). " width="100%" height="100%">
Journal: Disease Models & Mechanisms
Article Title: Validation of a model of rheumatoid arthritis using mice reconstituted with patient peripheral blood mononuclear cells
doi: 10.1242/dmm.052294
Figure Lengend Snippet: Human and mouse inflammatory markers in arthritic joints and plasma are increased in NSG-RA mice. Mice were treated as described in Fig. 1 . (A) RT-PCR expression analysis of IFNG , TNFA , Saa1 and Cxcl13 depicted as Cumming plots. CT values over 40 cycles and undetermined values were considered as not expressed and set to zero. (B) Plasma expression levels of human IFNγ, IL-17A, IL12p70 and TNFα detected by Luminex assay and displayed as Cumming plots. The upper part of the plot presents each data point in a swarmplot. The mean and s.d. of each group is plotted as a gapped line, where the vertical lines correspond to the mean±s.d. and the mean itself is depicted as a gap in the line (*** P ≤0.001, ** P ≤0.01, * P ≤0.05). Statistical analysis was performed with R (Kruskal–Wallis test). mRNA levels are depicted as lg(−deltaCT). RA, N =3, n =17; RA+, N =4, n =27; nonRA, N =1, n =6; nonRA+, N =1, n =6. Variations in sample sizes can be attributed to challenges encountered during RNA isolation and the exclusion of outliers during data processing. Values deviating more than sixfold from the mean were considered outliers. As indicated in the key to the right of the lower plot in B, plasma from the RA5 samples was not included in the human TNFα analysis, resulting in a decreased sample size for that plot. N =number of donors, n =total number of mice. Mice were reconstituted with PBMCs from patients with RA (RA3, n =6; RA4, n =6; RA5, n =11; RA7, n =6; RA8, n =15) and an unaffected individual (RA6, n =12).
Article Snippet: Single-tube TaqMan gene expression assays (Thermo Fisher Scientific) included the housekeeping gene Gapdh (Mm99999915_g1) and the markers TNFA (Hs01113624_g1), IFNG (Hs00989291_m1), Cxcl13 (Mm04214185_s1) and Saa1 (Mm00656927_g1).
Techniques: Clinical Proteomics, Reverse Transcription Polymerase Chain Reaction, Expressing, Luminex, Isolation
Fig. 7 . (A) Plasma levels of human TNFα, human IL12p70 and murine Cxcl9 presented as Cumming plots. Challenged, non-treated NSG-RA mice ( N =2, n =15 in total) were compared to challenged, infliximab-treated ( N =2, n =15 in total) or challenged, prednisolone-treated ( N =2, n =12 in total) mice. Outliers (values above 44,000 for Luminex) or undetermined values were removed for data analysis. (B) RT-PCR expression analysis of TNFA , Cxcl13 , Saa1 and IFNG calculated as lg(-deltaCT) and presented as Cumming plots. Challenged, non-treated NSG-RA mice ( N =3, n =21 in total) were compared to challenged, infliximab-treated ( N =3, n =15 in total) or challenged, prednisolone-treated ( N =2, n =11 in total) mice. The upper part of the plot presents each data point in a swarmplot. The mean and s.d. of each group is plotted as a gapped line, where the vertical lines correspond to the mean±s.d., and the mean itself is depicted as a gap in the line (*** P ≤0.001, * P ≤0.05). N =number of donors, n =total number of mice. Statistical analysis was performed with R (Mann–Whitney U -test). For raw data, see
Table S7 . Mice were reconstituted with PBMCs from patients with RA (RA5, n =12; RA7, n =18; RA8, n =18). " width="100%" height="100%">
Journal: Disease Models & Mechanisms
Article Title: Validation of a model of rheumatoid arthritis using mice reconstituted with patient peripheral blood mononuclear cells
doi: 10.1242/dmm.052294
Figure Lengend Snippet: Cytokine concentration in plasma and expression of human and mouse proinflammatory cytokines in arthritic joints are decreased in response to treatment with prednisolone or infliximab. Mice were treated as described in Fig. 7 . (A) Plasma levels of human TNFα, human IL12p70 and murine Cxcl9 presented as Cumming plots. Challenged, non-treated NSG-RA mice ( N =2, n =15 in total) were compared to challenged, infliximab-treated ( N =2, n =15 in total) or challenged, prednisolone-treated ( N =2, n =12 in total) mice. Outliers (values above 44,000 for Luminex) or undetermined values were removed for data analysis. (B) RT-PCR expression analysis of TNFA , Cxcl13 , Saa1 and IFNG calculated as lg(-deltaCT) and presented as Cumming plots. Challenged, non-treated NSG-RA mice ( N =3, n =21 in total) were compared to challenged, infliximab-treated ( N =3, n =15 in total) or challenged, prednisolone-treated ( N =2, n =11 in total) mice. The upper part of the plot presents each data point in a swarmplot. The mean and s.d. of each group is plotted as a gapped line, where the vertical lines correspond to the mean±s.d., and the mean itself is depicted as a gap in the line (*** P ≤0.001, * P ≤0.05). N =number of donors, n =total number of mice. Statistical analysis was performed with R (Mann–Whitney U -test). For raw data, see Table S7 . Mice were reconstituted with PBMCs from patients with RA (RA5, n =12; RA7, n =18; RA8, n =18).
Article Snippet: Single-tube TaqMan gene expression assays (Thermo Fisher Scientific) included the housekeeping gene Gapdh (Mm99999915_g1) and the markers TNFA (Hs01113624_g1), IFNG (Hs00989291_m1), Cxcl13 (Mm04214185_s1) and Saa1 (Mm00656927_g1).
Techniques: Concentration Assay, Clinical Proteomics, Expressing, Luminex, Reverse Transcription Polymerase Chain Reaction, MANN-WHITNEY
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