Skip to main content
. Author manuscript; available in PMC: 2018 Nov 13.
Published in final edited form as: Cancer Cell. 2017 Nov 13;32(5):654–668.e5. doi: 10.1016/j.ccell.2017.10.005

Figure 7. The mechanisms regulating inhibition of chemokines production by CSF1.

Figure 7

A. mRNA for Cxcl1 (by qPCR) (left) and Cxcl1 protein by ELISA (right) in mouse fibroblasts cultured for 6 days with CSF1 in the presence (during last three days of culture) of class I HDAC inhibitor entinostat. Mean and SD (n=3) are shown. ***-p<0.001 in Student’s t test from control. B–F. The effect of entinostat in vivo. LLC TB mice were treated with entinostat (10 mg/kg, p.o.) daily for 10 days starting from day 15 after tumor inoculation. Expression of mRNA for chemokines (qPCR) in tumor tissues. **-p<0.01 in Student’s t test from control (n=3) (B). Tumor cells were isolated at the end of the treatment and the amount of CSF1 was measured by ELISA (n=6) (C). CAF were sorted from tumor tissues and the amount of Cxcl1 was measured in cell lysates by ELISA. Mean and SD are shown. ****-p<0.0001 in Student’s t test from control (n=4) (D). CSF1R amount in CAF (E). The presence of PMN-MDSC and TAM in tumors of LLC (n=10) and CT-26 (n=3) TB mice treated with entinostat. Mean and SD are shown *-p<0.05; **-p<0.01 in Student’s t test from control (F). G. Mouse fibroblasts were treated with CSF1 for 4 days, followed by 2-day treatment of entinostat. ChIP of Cxcl1 promoter was performed using acetylated histone H3 or acetylated histone H4 antibody. The results are expressed as DNA enrichment, normalized to corresponding input values. Control IgG was used as a negative control (n=3). H. ChIP of Cxcl1 promoter was performed using HDAC2 specific antibody (n=3).