Fig. 3

MerTK ASO reshapes the population of immune cells in TME in favor of antitumor immune response A The percentages of MerTK⁺ tumor-associated macrophages (TAMs). B The MerTK expression level in TAMS. C M1/M2 ratio. D CD4⁺ T cell/CD45⁺ cell ratio. E CD8⁺ T cell/CD45⁺ cell ratio. F The percentages of GrB⁺ CD8⁺ T cells out of total CD8⁺ T cells. G Densities of CD8⁺, CD4⁺, NK, CD4⁺FOXP3⁺, GranzymB⁺CD8⁺, and GranzymB⁺NK cells in the secondary tumors stained with multiplex immune flouorescence. H Representative multiplex immune fluorescence images of Fig. G. The mice were treated with different combinations of XRT, MerTK ASO, αPD1, and αCTLA4, as indicated in Fig. 1A and Fig. 2A, and were sacrificed on day 21. Both the primary and the secondary tumors were harvested and stained with αCD45-PerCP-Cy5.5, αCD4-PE/Dazzle594, αCD8-FITC, αGrB-Pacific Blue, αGr1-BV510, αCD11b-APC Fire750, αF4/80-Alexa Fluor 700, αCD38-PE-Cy7, αCD206-PE, αMertK-APC. The secondary tumors tissues from groups Control, XRT, MerTK, XRT+αCTLA4, XRT+MerTK, XRT+MerTK+ αCTLA4 were also fixed and processed for multiplex immune fluoresce and further stained with DAPI, FOXP3, CD4, CD8, NKP46, and granzymB. All the statistics were compared with two-tailed t tests and expressed as mean value ± SEM. P values of <0.05 indicates statistical significance. *P<0.05, **P<0.01, ***P<0.001, NS denotes not significant