Journal: Journal of Immunotherapy (Hagerstown, Md. : 1997)

Article Title: NUSAP1 Promotes Immunity and Apoptosis by the SHCBP1/JAK2/STAT3 Phosphorylation Pathway to Induce Dendritic Cell Generation in Hepatocellular Carcinoma

doi: 10.1097/CJI.0000000000000531

Figure Lengend Snippet: NUSAP1 intervene in the differentiation of PBMCs around HCC into DC. (A) Elisa assay test IL-6 levels in the cell supernatant; (B and C) After co-culture, the expression levels of the surface factors CD1a and CD86 of PBMCs were determined by flow cytometry; (D and E) After LMT-28 intervene, the expression levels of the surface factors CD1a and CD86 of PBMCs were determined by flow cytometry; (F and G) After Coumermycin A1 intervene, the expression levels of the surface factors CD1a and CD86 of PBMCs were determined by flow cytometry; (H) After co-culture, WB detect Apoptosis-related proteins in HCC.

Article Snippet: According to the instructions of IL-6 ELISA kits purchased from Boster Biological Co. (Cat# MEK2004; Boster, USA), IL-6 level in the cultured cell supernatants of the different groups were detected, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Expressing, Flow Cytometry

Journal: Disease Models & Mechanisms

Article Title: Neurofibromin 1 mutations impair the function of human induced pluripotent stem cell-derived microglia

doi: 10.1242/dmm.049861

Figure Lengend Snippet: RNA sequencing reveals few differences between NF1 -mutant and CTL hiMGL cells, whereas the induced secretome remains unchanged. (A) Principal component analysis (PCA) plot generated from RNA sequencing data (CTL, blue; M1, red; M3, yellow). (B) Heatmap showing unsupervised hierarchical clustering generated from RNA sequencing data for CTL, M1 and M3 hiMGL cells. (C,D) Volcano plot demonstrating genes differentially expressed between M1 versus M3 (C) or between M1 and M3 versus CTL (D) (FDR<0.05, fold change cutoff of −5 and 5). Grey dots indicate genes with no change (N/C) in expression, blue dots indicate genes with decreased expression and red dots indicate genes with increased expression. (E) Relative mRNA expression (R.E.) of Toll-like receptor 4 ( TLR4 ) in CTL and NF1 -mutant (M1-M3) hiMGL cells by quantitative RT-PCR. Gene expression levels are shown relative to expression in CTL hiMGL cells. The housekeeping gene TBP was used for normalization ( n =3). Data were normalized to TLR4 expression levels in CTL hiMGL cells. Results are represented as the mean±s.e.m. Data were analyzed by one-way ANOVA. (F) Multiplex immunoassay was used to detect TNF-α (left) and IL-6 (right) in supernatants from CTL and NF1 -mutant (M1-M3) hiMGL cells in response to 1 μg/ml LPS stimulation for 24 h (n=3-4). Data between basal and LPS conditions were analyzed using two-tailed unpaired t -tests. Results are represented as the mean±s.e.m. Cytokine release under basal or LPS conditions was not significantly different between CTL and M1, M2 and M3 hiMGL cells (one-way ANOVA). The data showing the release of additional cytokines are included in Fig. S4 . n.s., not significant; * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: The MILLIPLEX MAP human cytokine/chemokine/growth factor panel A – immunology multiplex assay (HCYTA-60K, Merck) was used according to the manufacturer's recommendations.

Techniques: RNA Sequencing Assay, Mutagenesis, Generated, Expressing, Quantitative RT-PCR, Multiplex Assay, Two Tailed Test