Figure 5. Effect of therapeutic HIF-1α and HIF-2α ASO treatment on hepatic macrophage pool in DEN-induced HCC mice.

HCC was induced by weekly intraperitoneal DEN injection for 25 weeks. Control mice received weekly intraperitoneal 0.9% NaCl injection. DEN-treated mice were intraperitoneally injected with 20 mg/kg HIF-1α ASO, HIF-2α ASO or scrambled ASO twice per week, in a therapeutic setting. Control mice received scrambled ASO for the same duration of the experiment. Upper part: Percentage of CD11b⁺Ly6C^-F4/80⁺Tim4⁺ Kupffer cells, CD11b⁺Ly6C⁺F4/80⁺Tim4^- monocytes and CD11b⁺Ly6C^-F4/80⁺Tim4^- monocyte-derived macrophages in live CD45⁺ single cell gate following treatment, measured by flow cytometry. The upper and lower dashed lines represent mean ± SD of the control mice. Bars represent mean ± SD of different treatment groups of DEN-treated mice (n = 6–7 per treatment group). ^* p < 0.05, ^** p < 0.01, ^*** p < 0.001 and ^**** p < 0.0001. Lower part: Number of live CD45⁺CD11b⁺Ly6C^-F4/80⁺Tim4⁺ Kupffer cells, CD45⁺CD11b⁺Ly6C⁺F4/80⁺Tim4^- monocytes and CD45⁺CD11b⁺Ly6C^-F4/80⁺Tim4^- monocyte-derived macrophages per gram liver tissue following treatment, measured by flow cytometry. The upper and lower dashed lines represent mean ± SD of the control mice. Bars represent mean ± SD of different treatment groups of DEN-treated mice (n = 6–7 per treatment group). ^* p < 0.05, ^** p < 0.01, ^*** p < 0.001 and ^**** p < 0.0001.