Fig. 4.

HDAC11 suppression results in reduced lymph node growth but increased metastasis. a Relative expression of a set of down-regulated array genes after HDAC11 suppression by shRNA. Statistical significance was measured by ANOVA in comparison to the control shR samples. b Levels of immunoprecipitated promoter regions for IL-10, RRM2, E2F7, and E2F8 in human MDA-MB-231 cell lines expressing either control or HDAC11 ORFs when pulling down with either IgG control or HDAC11 antibodies. Statistical significance was measured by ANOVA. c Levels of immunoprecipitated promoter regions for RRM2, E2F7, and E2F8 in 4T1 cell lines stably expressing either control or HDAC11 shR when pulling down with either IgG control, acetyl-H3, or acetyl-H4 antibodies. Statistical significance was measured by ANOVA. d Western blots for E2F8 and RRM2 in 4T1 lines expressing control or HDAC11 shRs. e LN tumor weights at day 35 following axillary LN micro-injection. f Take rates of LN-micro-injected 4T1-shHDAC11 cell lines compared to 4T1-shCtrl cells. P value obtained using a χ² contingency test. g LN tumor volumes throughout the duration of the experiment (n = 13–14 mice/group). Statistical significance was measured by unpaired one-sided Student’s t tests. h Lung micro-metastasis enumeration for LN-micro-injected mice by mCh + flow cytometry. i Lung metastasis index after normalization to LN tumor size (n = 13–14 mice/group). Statistical significance was measured by unpaired one-sided Student’s t tests (h + i). j, k Disease-free survival curves from the BreastMark collection comparing patients with high and low levels of RRM2 in all available breast cancer samples (n = 2652 patients), as well as in LN+ cases (n = 744 patients). Statistical significance was measured by log-rank test. P values are indicated as *p < 0.05, **p < 0.01, and ***p < 0.001