Journal: Cell Death & Disease

Article Title: Chromatin accessibility analysis identifies the transcription factor ETV5 as a suppressor of adipose tissue macrophage activation in obesity

doi: 10.1038/s41419-021-04308-0

Figure Lengend Snippet: A BMDMs were transfected with siRNA negative control (siRNA NC) or siRNA specific to Etv5 (siRNA Etv5 ) 48 h and Il6 expression was assessed by RT-qPCR 48 h later. Il6 expression levels of Raw264.7 cells after knockdown of Etv5 using shRNA B or overexpression of Etv5 C . Il6 expression levels of Raw264.7 cells after knockdown of Etv5 using shRNA D or overexpression of Etv5 E under MMe induction condition. F Il6 expression levels of BMDMs after treatment with palmitate, glucose, and insulin alone or with a combination of these compounds for 24 h. G–I Measuring IL-6 protein levels in the supernatant of Raw264.7 cell culture by ELISA. Cells were treated with palmitate (P) or PGI G , or underwent Etv5 knockdown using siRNA H , or shRNA I . Data in this result were all representative of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001, Student’s t test.

Article Snippet: Supernatants were collected after macrophage culture and IL-6 protein concentrations were measured using a mouse IL-6 ELISA kit (E-MSEL-M0001, Elabscience, China) according to the manufactories’ instructions.

Techniques: Transfection, Negative Control, Expressing, Quantitative RT-PCR, Knockdown, shRNA, Over Expression, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cell communication and signaling : CCS

Article Title: Tumorous IRE1α facilitates CD8 + T cells-dependent anti-tumor immunity and improves immunotherapy efficacy in melanoma.

doi: 10.1186/s12964-024-01470-8

Figure Lengend Snippet: Fig. 4 Tumorous IRE1α promotes the secretion of Th1-related chemokine and cytokines by activating NF-κB. A, D Immunoblotting analysis of IRE1α, p-IRE1α, p65, p-p65, XBP1s and GAPDH in A2058 cells treated with TG (0.5 µM) or HA15 (10 µM) for 24 h after pretreated with or without STF-083010 (10 µM), MKC8866 (0.5 µM) or IRE1α siRNA for 24 h. B-C, E-H Relative mRNA level (n = 6) and ELISA (n = 4) analysis of CXCL9, CXCL10, CXCL11, TNF-α and IL-6 in A2058 cells treated with TG (0.5 µM) or HA15 (10 µM) for 24 h after pretreated with or without STF-083010 (10 µM), MKC8866 (0.5 µM) or BAY 11-7085 (1 µM) for 24 h. I A2058 cells treated with TG (0.5 µM) or HA15 (10 µM) for 24 h after pretreated with or without MKC8866 (0.5 µM) for 24 h were subjected to ChIP with normal mouse IgG, NF-κB or Pol-II antibody as indicated (n = 3). J A2058 cells treated with TG (0.5 µM) or HA15 (10 µM) for 24 h were subjected to ChIP with normal mouse IgG, XBP1s or Pol-II antibody as indicated (n = 3). ChIP samples were analyzed by qPCR using primers indicated in Additional file 2: Table S3. Data are representative of at least three independent experiments and shown as mean ± SD. Two-tailed Student’s t-test or two-way ANOVA (*p < 0.05; **p < 0.01; ***p < 0.001)

Article Snippet: ELISA analysis on culture medium of melanoma cells or the blood serum of mice after indicated treatment was performed using the Human IL-6 ELISA Kit (Neobioscience, EHC007.96), Human TNF-α ELISA Kit (Neobioscience, EHC103a.96), Human MIG/CXCL9 ELISA Kit (Neobioscience, EHC114.96), Human IP-10/CXCL10 ELISA Kit (Neobioscience, EHC157.96), Human I-TAC/ CXCL11 ELISA Kit (Neobioscience, EHC084.96), Mouse TNF-α ELISA Kit (Elabscience, E-MSEL-M0002), Mouse IL-2 ELISA Kit (Elabscience, E-MSEL-M0036), Mouse IL-6 ELISA Kit (Elabscience, E-EL-M0044c), Mouse ELISA Kit MIG/CXCL9 ELISA Kit (Elabscience, E-ELM3077), Mouse IP-10/CXCL10 ELISA Kit (Elabscience, E-EL-M0021c), Mouse I-TAC/CXCL11 ELISA Kit (Elabscience, E-EL-M0056c) and Mouse HMGB-1 ELISA Kit (Elabscience, E-EL-M0676c) according to the manufacturer’s instructions.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test