Fig 1: Higher levels of CXCL9 but not of IFN? in serum of WT compared to Mcpt6-/- mice. 50,000 B16F10 cells were injected subcutaneously in the hip region of WT or Mcpt6-/- mice. After termination of the experiment, blood was collected. Blood was also collected from naïve WT and Mcpt6-/- mice. (a-b) Levels of CXCL9 in serum from (a) B16F10-injected (n = 12) and (b) naïve (n = 11) mice, measured by ELISA. Note the higher levels of CXCL9 in serum from B16F10-injected WT versus Mcpt6-/- mice, whereas no differences in CXCL9 levels were seen between WT and Mcpt6-/- naïve mice. (c) IFN? levels in serum from B16F10-injected WT and Mcpt6-/- mice, measured by ELISA (n = 12). Results are presented as mean values ± SEM; Mann–Whitney test. *p = .05
Fig 2: APCs expressing CXCL10 administered intravenously with LIPU-induced BBBD were associated with a significant increase in immunocompetent mouse survival durations. A, Schema of the lentivirus gene transfer plasmid structure for murine CXCL9 or CXCL10. B, Gene transfer plasmid of control methionine-deficient green fluorescent protein alone. C, Flow cytometry analysis of the percentage of CD3+ T cells within the total CD45+ immune cell infiltration within intracerebral gliomas in the different treatment groups (n = 5–7/group). Statistics: PBS versus CXCL10 with ultrasound, P = 0.0167; CXCL10 versus CXCL10 with ultrasound, P = 0.0412; PBS with ultrasound versus CXCL10 with ultrasound, P = 0.0004. D, Flow cytometry analysis of CD4+ T cells within the total CD45+ immune cell infiltration within intracerebral gliomas. Statistics: PBS versus CXCL10 with ultrasound, P = 0.0098; CXCL10 versus CXCL10 with ultrasound, P = 0.0371; PBS with ultrasound versus CXCL10 with ultrasound, P = 0.0003. E, Flow cytometry analysis of CD8+ T cells within the total CD45+ immune cell infiltration within intracerebral gliomas. Statistics: PBS with ultrasound versus CXCL10 with ultrasound, P = 0.0023; PBS versus CXCL10 with ultrasound, P = 0.153; CXCL10 versus CXCL10 with ultrasound, P = 0.2154, as assessed by the two-sided unpaired t test. F, Treatment schema for C57BL/6 mice bearing GL261 tumors treated with CXCL9 or CXCL10 APCs. G, Kaplan–Meier survival analysis of mice treated with PBS, i.c. CXCL10 APCs, i.v. CXCL10 APCs, or i.v. CXCL10 APCs with ultrasound. PBS with sonication control for the GL261 model is demonstrated in Supplementary Fig. S1A. The MS durations in the treatment groups were as follows: PBS (3 mice), 24 days; i.c. CXCL10 APCs (4 mice), 28 days; i.v. CXCL10 APCs (4 mice), 24 days; i.v. CXCL10 APCs with ultrasound (3 mice), 34 days. Statistics: PBS versus i.c. CXCL10 APCs, P = 0.0476; PBS versus i.v. CXCL10 APCs, P = 0.6041; PBS versus i.v. CXCL10 APCs with ultrasound, P = 0.0246; i.c. CXCL10 APCs versus i.v. CXCL10 APCs, P = 0.6349; i.c. CXCL10 APCs versus i.v. CXCL10 APCs with ultrasound, P = 0.0213; i.v. CXCL10 APCs versus i.v. CXCL10 APCs with ultrasound, P = 0.0415.
Fig 3: T cells and monocytes contribute to hypercytokinemia in the R688*/R688* proband. a Serum concentration of the cytokines IL-1ß, IL-1RA, IL-6, IL-10, IL-17A, IL-18, IFN?, CXCL9, and TNF in HCs (n = 4) and proband (two biological replicates) or HCs (n = 3) and proband (one biological replicate) in the case of the cytokine IL-17A. Mean and SEM are depicted. b, c ELISA of TNF and IFN? produced by in vitro PMA/ionomycin stimulated CD4+ T cells (b) or CD8+ T cells (c) of HCs (n = 4) and proband. d ELISA of TNF produced by monocytes of HCs (N = 4) or proband treated in vitro overnight with LPS. e RT-qPCR quantifying TNF and IFNG transcripts in PMA/ionomycin stimulated T cells in absence or presence of mepazine pretreatment (20'). Cells were sampled 1 h after stimulation. Data was normalized using the housekeeping genes HPRT and GAPDH. HCs (n = 6). *p < 0.05 (paired t-test). R688* proband (n = 2). Data shown are accumulated from two independent experiments (a, e) or representative for two independent experiments (b–d). Source data are provided as a Source Data file
Fig 4: Detection of inflammation and oxidative-reduction related factors in heart tissue of mice using ELISA method (A) iNOS, (B) COX-2, (C) CCL9, (D) CXCL1, (E) CXCL9 and (F) CXCL11 in mouse heart tissues. Data are presented as the mean ± SD. Each experiment was repeated three times independently. *P<0.05 vs. NC group. #P<0.05 vs. OT group. DHE, dehydrocostus lactone; TXNIP, thioredoxin-interacting protein; NC, negative control; OT, DHE treatment group; OI, DHE treatment combined with TXNIP inhibition group; OH, DHE treatment combined with TXNIP overexpression group; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; CCL9, C-C motif ligand 9; CXCL, chemokine C-X-C motif ligand.
Fig 5: Mcpt6 deficiency affects gene expression in melanoma tumors. Total RNA was isolated from tumors of WT and Mcpt6-deficient mice. Subsequently, total RNA was subjected to qPCR analysis of expression of Cxcl9, Tgtp2, Gbp10, Gbp2, and miR3098. Expression of genes was evaluated relative to either GAPDH (protein-encoding genes) or 5S-rRNA (miRNAs), and normalized to WT mice. Note the increased expression of Cxcl9, Tgtp2, and Gbp10, and lower expression of miR3098 in tumors from WT versus Mcpt6-/- mice. Results are presented as mean values ± SEM (n = 7–13); Mann–Whitney test. *p = .05
Supplier Page from Abcam for Mouse CXCL9 ELISA Kit