Supplementary Figures S1, S2, S3 from Radiation Followed by OX40 Stimulation Drives Local and Abscopal Antitumor Effects in an Anti–PD1-Resistant Lung Tumor Model

Supplementary Figure S1. XRT (12 Gyx 2) with α-OX40 treatment effect on (A) tumor growth and (B) lung metastases in induction and adjuvant settings. (C) A representative picture depicting the decrease in lung tumor nodules with the combination therapy after staining with Bouin's fixative solution. /// Supplementary Figure S2. Effect of radiation (XRT) on immune-cell profiles in splenocytes (Spl) and draining lymph nodes (dLN). Representative flow cytometry gating of (CD4+OX40+) lymphocytes isolated from (A) spleen and (B) tumor draining lymph node of mice at 48 h after XRT (12 Gyx 3) or no XRT to anti-PD1 resistant tumors. /// Supplementary Figure S3. XRT upregulates OX40+ CD8 T-cells and enhances OX40L expression in neutrophils. (A) In anti-PD1-resistant 344SQ tumor model, XRT given as three 12-Gy fractions (total dose 36 Gy) increased the percentages of CD8+ T-cells expressing OX40 in the splenocytes 48 h after last dose of radiation. (B) Percent neutrophils expressing OX40L isolated from splenocytes 48 h after XRT. Cells are first gated on leukocytes then Gr1hi CD11b+ cells then on OX40L+ population. *P {less than or equal to} 0.05 is considered significant.