(A) Scheme for experimental design to investigate the immunomodulatory effect of RFA in TdLN and cLN. (B) Representative photomicrographs and quantification of TRITC-labeled naïve OT-I CD8+ T cells (red) in cLN and TdLN of C57BL/6 mice bearing B16-OVA tumors after sham procedure or RFA. LN were counterstained for PNAd to identify cuboidal endothelium lining HEV (green). (C, D) In vivo short-term (1 hour, left; 12 hour, right) homing of TRITC-labeled naïve (C) or activated (D) Clone 4 CD8+ T cells in TdLN and cLN from BALB/c mice bearing CT26-HA tumors after sham procedure or RFA. Data are the mean ± SEM of ≥ 10 fields for non-sequential cyrosections analyzed from independent mice (n=3 mice per group) and are representative of 2 experiments. ***, P < 0.001 by unpaired, two-tailed Student’s t-test. Scale bars (B-D), 100 μm.