Fig 1: Effects of TCDD on expression and secretion of chemokines in astrocytes. Cells were incubated with different concentrations of TCDD (0.03, 0.01, 0.03, 0.1, or 0.3 nmol/L) or 0.1% DMSO for 24 h. The mRNA expression levels of (A) Cxcl2, (B) Ccl7, and (C) Cxcl10 were determined by qPCR in dose-dependent effects of TCDD for 24 h. Cells were treated with 0.03 nmol/L TCDD or 0.1% DMSO for 12, 24, 36, 48, 72, or 96 h. The mRNA expression levels of (D) Cxcl2, (E) Ccl7, and (F) Cxcl10 were determined by qPCR in TCDD time-dependent effects. GAPDH was an internal control for quantification. The protein expression of (G) Cxcl2, (H) Ccl7, and (I) Cxcl10 were determined by ELISA. Data are shown as fold of control and expressed as mean ± SEM (n = 3) and each independent sample was detected in triplicate. Statistical analysis was done by one-way ANOVA with Bonferroni test in dose-dependent effects of TCDD for 24 h or two-way ANOVA with Bonferroni test in TCDD time-dependent effects. Statistically significant difference *p < 0.05, **p < 0.01, and ***p < 0.001 compared with the DMSO groups.
Fig 2: Effects of FICZ on the expression of AhR downstream genes and chemokines in astrocytes. After 3, 6, 9, 12, 24, or 30 h treatment with FICZ (50 nmol/L) or 0.1% DMSO. The mRNA level of (A) CYP1A1, (C) AHRR, (D) Cxcl2, (E) Ccl7, and (F) Cxcl10 were determined via qPCR. GAPDH was used as an internal control. (B) CYP1A1 activity of astrocytes was measured by P450-Glo Assays. Values were shown as fold of control (DMSO group) and expressed as mean ± SEM (n = 3) and each independent sample was detected in triplicate. Statistical analysis was done by two-way ANOVA with Bonferroni test. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with control.
Fig 3: Roles of AhR pathway in the effects of TCDD on autonomous motility and the expression of chemokines of the astrocytes. Astrocytes treated with TCDD (0.03 nmol/L) or 0.1% solvent control together with or without CH223191 (1 μmol/L) were continuously tracked for 48 h. (A) Displacement of cell motility was collected separately at 12, 24, 36, and 48 h. Values were expressed as mean ± SEM (n ≥ 208) and each independent sample was detected in sextuple. (B) Cxcl10, Cxcl2 and Ccl7 mRNA level in cultured astrocytes were inhibited by CH223191 incubated with 0.03 nmol/L TCDD or 0.1% DMSO for 24 h. Data were shown as fold of control and expressed as mean ± SEM (n = 3) and each independent sample was detected in triplicate. Statistical analysis was done by two-way ANOVA with Bonferroni test. *p < 0.05, **p < 0.01, and ***p < 0.001 compared with control.
Supplier Page from Abcam for Rat CINC-3 ELISA Kit (CXCL2)