Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IFN-beta1a inhibits the secretion of Th17-polarizing cytokines in human dendritic cells via TLR7 up-regulation.

doi: 10.4049/jimmunol.0802226

Figure Lengend Snippet: FIGURE 2. IFN-1a treatment up-regulates TLR7 and its signaling pathway, while it down-regulates the expression of IL-1R and IL-1/. A, Three 106 DCs per condition derived from 10 RR MS patients were pretreated with IFN-1a for 24 h and then matured with LPS for 2 h before RNA extraction. TLR7, MyD88, IRAK4, TRAF6, IL-1R1, IL-1, and IL-1 gene expression was measured by RT- PCR. The results are presented as relative gene expression normalized for the 18S RNA expression. B, Three 106 DCs per condition from 16 RR MS patients were pretreated with IFN-1a for 24 h and then matured with LPS for 48 h. DCs were stained with PE-Cy-CD11c, allophycocyanin-CD1a mAb, and anti-IL-1R1 Ab, followed by FITC-conjugated secondary Ab, anti-TLR7 primary Ab, and FITC-conjugated secondary Ab. FITC-conjugated IgG isotype Ab was used as a control. The percentage of positive cells was determined in the CD1a-gated population. C, Three 106 DCs per condition derived from three RR MS patients were pretreated by IFN-1a for 24 h and then matured with LPS for 5 h before protein extraction. The expression of TLR7, MyD88, IRAK4, TRAF6, IL-1R1, and -actin were measured by Western blotting. The results present one of three similar experiments.

Article Snippet: Nonspecific binding was blocked for 1 h in the blocking buffer (10 mM Tris (pH 7.5), 150 mM NaCl, 2% fish gelatin, and 1% OVA in double-distilled water) and incubated overnight with rabbit anti-human Ab for TLR7 (Abcam), MyD88, TRAF6 (Santa Cruz Biotechnology), and IRAK4 (ProSci) and with mouse anti-human Ab for IL1R1 and -actin (Santa Cruz Biotechnology).

Techniques: Expressing, Derivative Assay, RNA Extraction, Gene Expression, Reverse Transcription Polymerase Chain Reaction, RNA Expression, Staining, Control, Protein Extraction, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IFN-beta1a inhibits the secretion of Th17-polarizing cytokines in human dendritic cells via TLR7 up-regulation.

doi: 10.4049/jimmunol.0802226

Figure Lengend Snippet: FIGURE 4. IFN-1a modifies MyD88/IRAK4/ TRAF6 signaling and cytokine production in DCs through its induction of TLR7 expression. Three 106/well DCs generated from six RR MS patients were plated in antibiotic-free medium. After a 24-h incubation at 37oC, the DCs were transfected with TLR7 siRNA or control siRNA. The siRNA-treated cells were harvested and cultured in serum-free me- dium in the absence or presence of IFN-1 for 24 h, followed by LPS maturation for 5 h before the pro- tein extraction, and for 48 h before the SN collec- tion. A, The expression of TLR7, MyD88, IRAK4, TRAF6, IL-1R1, and -actin was measured by Western blotting. The results present one of three similar experiments. B, The production of IL-1, TGF-1, IL-23/p19, and IL-27 was measured by ELISA. The results present six experiments per- formed in triplicate. Statistical analysis was per- formed using a repeated measures ANOVA. , p 0.05; , p 0.01; and , p 0.001.

Article Snippet: Nonspecific binding was blocked for 1 h in the blocking buffer (10 mM Tris (pH 7.5), 150 mM NaCl, 2% fish gelatin, and 1% OVA in double-distilled water) and incubated overnight with rabbit anti-human Ab for TLR7 (Abcam), MyD88, TRAF6 (Santa Cruz Biotechnology), and IRAK4 (ProSci) and with mouse anti-human Ab for IL1R1 and -actin (Santa Cruz Biotechnology).

Techniques: Expressing, Generated, Incubation, Transfection, Control, Cell Culture, Extraction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: A novel immune cell signature for predicting osteosarcoma prognosis and guiding therapy

doi: 10.3389/fimmu.2022.1017120

Figure Lengend Snippet: Construction of a risk model. (A) Univariate COX regression analysis for the 94 genes associated with the infiltration of resting dendritic cells. (B, C) LASSO analysis for other important genes associated with the survival rate of OS. (D) The HR and p value of genes (including AOC3, CDK6, COL22A1 and RNASE6) under multivariate COX analysis are shown.

Article Snippet: The primary antibodies used were AOC3 (1:500; Cat no. 66834-1-Ig, Proteintech, Wuhan, China), CDK6 (1:200; Cat no. 14052-1-AP, Proteintech, Wuhan, China), COL22A1 (1:250; Cat no. ab121846; Abcam, USA), and RNASE6 (1:100; Cat. ab121111; Abcam, USA), for 14 hours at 4°C.

Techniques:

Journal: Frontiers in Immunology

Article Title: A novel immune cell signature for predicting osteosarcoma prognosis and guiding therapy

doi: 10.3389/fimmu.2022.1017120

Figure Lengend Snippet: The risk model exhibits high prognostic value in the TARGET discovery cohort. (A) OS tissues in TARGET were divided into low- and high-risk groups, according to median risk scores. (B) Kaplan survival analysis indicated the difference between low- and high-risk group OS tissues in TARGET. (C–E) ROC analyses of the risk model for the 1-year, 3-year, and 5-year survival rates for OS patients in TARGET. (F) Alive and death cases between low- and high-risk group OS tissues in TARGET. (G) Expression of COL22A1, CDK6, RNASE6 and AOC3 between low- and high-risk group OS tissues in TARGET.

Article Snippet: The primary antibodies used were AOC3 (1:500; Cat no. 66834-1-Ig, Proteintech, Wuhan, China), CDK6 (1:200; Cat no. 14052-1-AP, Proteintech, Wuhan, China), COL22A1 (1:250; Cat no. ab121846; Abcam, USA), and RNASE6 (1:100; Cat. ab121111; Abcam, USA), for 14 hours at 4°C.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: A novel immune cell signature for predicting osteosarcoma prognosis and guiding therapy

doi: 10.3389/fimmu.2022.1017120

Figure Lengend Snippet: The risk model exhibits high prognostic value in the GSE21257 verification cohort. (A) OS tissues in GSE21257 were divided into low- and high-risk groups according to median risk scores. (B) Kaplan survival analysis indicated the difference between low- and high-risk group OS tissues in GSE21257. (C–E) ROC analysis of the risk model for the 1-year, 3-year, and 5-year survival rates for OS patients in GSE21257. (F) Alive and death cases between low- and high-risk group OS tissues in GSE21257. (G) Expression of COL22A1, CDK6, RNASE6 and AOC3 between low- and high-risk group OS tissues in GSE21257.

Article Snippet: The primary antibodies used were AOC3 (1:500; Cat no. 66834-1-Ig, Proteintech, Wuhan, China), CDK6 (1:200; Cat no. 14052-1-AP, Proteintech, Wuhan, China), COL22A1 (1:250; Cat no. ab121846; Abcam, USA), and RNASE6 (1:100; Cat. ab121111; Abcam, USA), for 14 hours at 4°C.

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: A novel immune cell signature for predicting osteosarcoma prognosis and guiding therapy

doi: 10.3389/fimmu.2022.1017120

Figure Lengend Snippet: The risk model has the potential to predict metastasis in patients with OS. (A) Non-metastasis and metastasis cases between low- and high-risk group OS tissues in the TARGET and GSE21257 cohorts. (B) ROC analysis for the diagnostic value of the risk model in the prediction of OS tissue metastasis. (C, D) IHC was performed to detect the expression of COL22A1, CDK6, RNASE6 and AOC3 in non-metastasis and metastasis OS tissues (magnification 200× and 400×). *p < 0.05; **p < 0.01; ns, no significant.

Article Snippet: The primary antibodies used were AOC3 (1:500; Cat no. 66834-1-Ig, Proteintech, Wuhan, China), CDK6 (1:200; Cat no. 14052-1-AP, Proteintech, Wuhan, China), COL22A1 (1:250; Cat no. ab121846; Abcam, USA), and RNASE6 (1:100; Cat. ab121111; Abcam, USA), for 14 hours at 4°C.

Techniques: Diagnostic Assay, Expressing