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. 2015 Feb 25;4(2):e978709. doi: 10.4161/2162402X.2014.978709

Figure 4.

Figure 4

(See previous page). Temozolomide treatment does not induce T-cell infiltration into skin tumors from RETAAD mice despite increased CXCL9 and CXCL10 expression. Cutaneous tumor-bearing RETAAD mice were treated with Temazolomide (TMZ) or dimethyl sulfoxide (DMSO) vehicle. (A) qRT-PCR analysis of gene expression of CD3, CD4 and CD8 in skin tumors following Temozolomide (TMZ) treatment. (B) Flow cytometry analysis of T cells in skin tumors from RETAAD mice at day 10 after TMZ treatment. (C) Immunofluorescence imaging to detect T cells in skin tumors at day 10 after TMZ treatment. (D) CXCL9 and CXCL10 gene expression in skin tumors following TMZ treatment. (E) Immunofluorescence imaging was performed to examine CXCL9 and CXCL10 protein expression in skin tumors at day 10 after TMZ treatment. (F) MFI quantification of CXCL9 and CXCL10 staining in skin tumors at day 10 after TMZ treatment. Data from panels: (A and D) are pooled from 3-5 mice per group; (B) are pooled from 2 separate experiments with 3-4 mice per group in each experiment (n = 6-8/group). Data from (F) are from 4-6 mice per group. Bars represent mean ± SD. Statistical analyses were performed using one-way ANOVA test with Bonferroni's post-test analysis in (A and D) and the unpaired two-tailed Student's t-test in (B and F); *p<0.05, **p<0.01. Images from (C and E) are representative of 5 independent experiments (n = 5 in each group) and scale bars in represent 200 μm.