Fig. 5: Chronic circadian disruption attenuates immune infiltration and creates a pro-tumour immune microenvironment.

a Percentage of tumour-infiltrating immune cells (TIC) in LD (n = 13) and JL (n = 13) tumours. Flow cytometric analysis revealed a lower percentage of TICs in JL tumours. Data are presented as a scatter dot plot with lines indicating the median with interquartile range (error bars). P-value obtained from an unpaired two-sided t-test. b Relative distribution of main immune cell types in TICs from LD and JL tumours. Data presented as pie charts displaying the mean values of 13 mice. c, d Tumour-associated macrophage (TAM) phenotypes in LD (n = 13) and JL (n = 13) tumours. Flow cytometry of JL tumours showed a significant reduction in the anti-tumour CD11b⁺MHC II^hi phenotype, with a significant increase in pro-tumour CD11b⁺MHC II^low TAMs. e Tumour-infiltrating lymphocytes (TIL) in LD (n = 10) and JL (n = 8) tumours. TILs are presented as percentage of CD3⁺ immune cells. The proportion of CD8+ TILs was significantly lower in JL tumours, resulting in an increase in the CD4/CD8 ratio. f Tumour-infiltrating CD4⁺FoxP3⁺ T cells in LD (n = 10) and JL (n = 8) mice. Flow cytometry revealed a significant increase of Treg and Treg/CD8⁺ ratio in primary tumours of JL mice. g CD4/CD8 ratio in the peripheral blood of LD (n = 8) and JL (n = 6) mice. c–g Data are presented as scatter dot plots with lines indicating the median with interquartile range (error bars). P-values obtained from unpaired two-sided t-test. Indicated (n) represent number of independent experiments as biological replicates.