Figure S2. CCL2 mRNA and protein level is detectable GL261 model of GBM. from CCL2 Produced by the Glioma Microenvironment Is Essential for the Recruitment of Regulatory T Cells and Myeloid-Derived Suppressor Cells

<p>(A) GL261 cells were orthotopically implanted into the right hemisphere of syngeneic immunocompetent C57BL/6 mice. At 1 week post-intracranial (i.c.) injection whole brains were homogenized followed by Percoll gradient separation of leukocytes and non-leukocytes. qRT-PCR was performed for CCL2 in mice intracranially-injected with GL261 cells (n = 4) or PBS-injected controls (n = 3). (B) Detection of CCL2 protein in both brain hemispheres at 1 week-i.c. GL261 (n = 3). (C) Immunoreactivity for CCL2 was detected at 1 week post-GL261 injection. 20Ã- and 63Ã- magnification. Scale bar = 50 microm. (D) qRT-PCR for CCL1, CCL28, CCL2, CCL22, CCL17, and CCL20 from non-leukocytes (NL) and leukocytes (L) as in (A). Images in C are representative of 3 independent replicates. Data are representative of at least 2 independent experiments. Data are represented as mean {plus minus} SEM; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by one-way ANOVA with Tukey's multiple comparisons test (A, D) or Student's t test (B).</p>