m cxcl10 ip 10 goat igg  (R&D Systems)

Journal: Cell Reports Medicine

Article Title: Histone methyl-transferase G9a inhibition boosts the efficacy of immune checkpoint inhibitors in experimental hepatocellular carcinoma

doi: 10.1016/j.xcrm.2026.102717

Figure Lengend Snippet: G9a inhibition potentiates the immunogenic effects of IFN-γ in HCC cells (A) Effect of CM272 and EZM8266 on IFN-γ-triggered CXCL10 production in murine (PM299L) and human (HuH7) HCC cells. Cells were treated with CM272 for 48 h or with CM272 for 24 h and then with IFN-γ (75 U/mL) for another 24 h or with IFN-γ alone for 24 h. PM299L were treated with 400 nM, and HuH7 received 1 μM of CM272 ( n = 3). For EZM8266, cells were pretreated for 48 h with EZM8266 (5 mM) and then with IFN-γ (75 U/mL) for another 24 h, as indicated. CXCL10 protein levels were measured by ELISA in cells’ conditioned media ( n = 3). (B) Effect of G9a inhibition with CM272 or EZM8266 on the expression of MHC class I complex protein (MHC-I) on the surface of PM299L cells. Cells were treated with IFN-γ and CM272 or EZM8266 as indicated in (A), and MHC-I levels were determined by FACS analysis ( n = 3). (C) ChIP analyses of H3K9me2 levels in the proximal promoter regions of CXCL10 and HLA-A genes in HuH7 cells treated with IFN-γ (75 U/mL) and CM272 (400 nM), as indicated in (A) ( n = 3). (D) Evaluation of the expression of transposable elements (TEs) and endogenous retroviral sequences (ERVs) by RNA-seq in NM53 murine HCC cells treated with IFN-γ, CM272, and their combination as indicated in (A). (E) Immunofluorescence analyses of dsRNA in PM299L HCC cells treated with CM272 (24 h) or EZM8266 (48 h). Representative images are shown. Scale bars, 10 μm. Right panel shows a control without primary antibody ( n = 3). Data are represented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. All the replicates represent biological replicates.

Article Snippet: CXCL9 and CXCL10 concentrations were measured in culture supernatants collected at the end of the incubation periods using commercial ELISA kits for mouse CXCL9 (DY492) and CXCL10 (DY466), both from R&D Systems, and an ELISA kit for human CXCL10 (550926) from BD Biosciences (Franklin Lanes, NJ, USA).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Expressing, Retroviral, RNA Sequencing, Immunofluorescence, Control

Journal: Journal of Advanced Research

Article Title: The pro-cancer and immunosuppressive activity of macrophage-transformed cancer-associated fibroblasts in oral squamous cell carcinoma

doi: 10.1016/j.jare.2025.07.027

Figure Lengend Snippet: MMTs recruit a large number of MDSCs through CXCL10 pathway. A Significantly upregulated genes in MMTs versus non-MMT fibroblasts from OSCC database (GSA-Human: HRA007439). B Differences in CXCL10 expression between non-MMT fibroblasts and MMTs were demonstrated in HNSCC (figure S1). C CXCL10 expression and release were detected by PCR and ELISA. D-H Exogenous CXCL10 was added to BMDMs, while the CXCL10 inhibitor AMG487 was administered to MMTs, followed by co-culturing with MDSCs. The migration of MDSCs was evaluated using transwell chambers and crystal violet staining ( D ). Flow cytometry was used to detect cell apoptosis ( E ), macrophage markers (F4/80 + CD11b + ) ( F ), as well as CFSE for assessing T cell proliferation inhibition ( G ). PCR and ELISA were employed to measure the expression of immunosuppressive factors IL10 and iNOS ( H ). I, J Tumor tissue photographs ( I ) and the tumor growth curve were taken and subjected to statistical analysis regarding volume and weight ( J ). K-M Flow cytometry was used to detect the proportion of MDSCs in the spleen ( K ) and tumors ( L ) of each group, while immunofluorescence staining ( M ) revealed the densest field of MDSCs within the tumor tissues across all groups. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs experimental group. “ns” represents no statistical significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: According to the manufacturer's instructions, the cytokine concentrations of TGFβ1, iNOS, IL-10, and CXCL10 in the supernatant were determined using ELISA kits (Boster, California, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Migration, Staining, Flow Cytometry, Inhibition, Immunofluorescence