Fig. 2.

Tumor inhibition and immune modulation by PARP1 AsiC and PARP1 inhibitor olaparib. (A) Growth of 4T1E orthotopic tumors in mice treated beginning on day 3 with EpCAM aptamer or PARP1 EpCAM-AsiC (5 mg/kg, every third day) or olaparib (50 mg/kg daily). (B to D) Flow cytometry analysis of TIL, harvested on day 14 after implantation from mice treated as in A, for (B) CD8⁺/CD4⁺Foxp3⁺ Treg and percentage of CD8⁺ (C) and CD4⁺ (D) TIL producing IFN-γ and TNF-α after PMA and ionomycin stimulation. (n = 4 in A–D). (E and F) Flow cytometry analysis of unrepaired DNA damage in CD45⁻EpCAM⁺ tumor cells as measured by mean fluorescence intensity (MFI) of γ-H2AX (E) and percentage of TUNEL⁺ tumor cells (F) in each group of day 14 tumors. (G) Relative mRNA expression by qRT-PCR of type I interferon genes in each group of day 14 tumors. (E–G) EpCAM aptamer: n = 4, PARP1 AsiC and olaparib: n = 5. Data shown are mean + SEM and are representative of two experiments. Statistical tests: (A) tumor growth curves were compared by calculating the area under the curve values for each sample followed by one-way ANOVA with Holm–Sidak’s multiple comparisons. (B–E) One-way ANOVA with Holm–Sidak’s multiple comparisons. (F) Kruskal–Wallis test with Dunn’s multiple comparisons. (G) Two-way ANOVA with Holm–Sidak’s multiple comparisons. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.