Fig. 3

RNA-sequencing analysis, hypoxia modulation and systematic immune response activation of local ¹³¹I-MnO₂-BSA treatment in 4T1-bearing mice model. a Volcano plots showing the DEGs between the control and ¹³¹I-MnO₂-BSA groups; b Heatmap of DEGs between the control and ¹³¹I-MnO₂-BSA groups; c Biological process analysis; d Cellular component analysis; e Molecular function analysis. f The Log2 fold change of DEGs related to immune and hypoxia response; g The p value of DEGs related to immune and hypoxia response; h Protein–protein interaction networks. G1 and G4 represent Control, ¹³¹I-MnO₂-BSA, respectively. i Immunofluorescence images of tumors slices in different groups, and tumor slices were harvested from those mice experiencing corresponding treatment in different groups, and then stained with anti-HIF-α, anti-CD8, anti-Foxp3, anti-F4/80, and anti-CD206 antibodies, respectively. j–o Statistical data of HIF-1α+ (j), CTLs+ (k), Foxp3+ (Treg, l), CTLs/Treg (m), F4/80+ (n) and CD206+ (o) areas in tumors after various treatments, which were obtained from i. The ratios were normalized in relation to the PBS group. P values were calculated by ANOVA (***p < 0.001; *p < 0.05; ns, not significant). G1–G4 represent Control, MnO₂-BSA, free ¹³¹I and ¹³¹I-MnO₂-BSA treatment, respectively. Data are expressed as mean ± SD (n = 3), dose: 500 µCi