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. Author manuscript; available in PMC: 2018 Jul 1.
Published in final edited form as: Hepatology. 2017 May 22;66(1):182–197. doi: 10.1002/hep.29184

Figure 5. DEN-induced HCC tumors in Alb-HMGB1−/− mice show decreased mitochondrial biogenesis and decreased adaptation to hypoxia.

Figure 5

(A and B) DEN-induced HCC tumors exhibit increased expression of PGC-1α compared to background liver. (C) PGC-1α, NRF1 and TFAM protein tumor levels measured in HMGB1 control and in Alb-HMGB1−/− mice showing decreased expression in tumors lacking HMGB1. (D) There was a significant decreased in mtDNA copy numbers measured by RT-PCR in tumors lacking HMGB1. (T, Tumor; NT, Non-tumor). ***p<0.01, ****p<0.001, ns: not significant. (E) Confocal microscopy shows decreased mitochondrial density (TOM20 levels) in HMGB1 deficient tumors. The median normalized TOM20+ area in control non-tumors was 0.025 (Range 0.001–0.038) versus 0.384 (Range 0.261–0.474) in control tumors. When looking at Alb-HMGB1−/− tumors, the median normalized TOM20+ area in Alb-HMGB1−/− non-tumors was 0.019 (Range 0.0004–0.056) versus 0.073 (Range 0.0053–0.192) in Alb-HMGB1−/− tumors. Representative magnified (60X–1.75z) images included of both control and Alb-HMGB1−/− tumors (Scale Bars 50μm). (T, Tumor; NT, Non-tumor). *p<0.05, **p<0.01, ns: not significant. Scale Bars 100μm, Nuclei (blue), Actin (green), TOM20(red). The values were based on analysis of tumors from 6 mice for each group. (F) There is marked increase in oxidative stress and formation of 4-HNE in Alb-HMGB1−/− tumors compared to HMGB1 control tumors. Median (Range) 4-HNE area (μm2) normalized by actin area (μm2): 0.046 (0.002–0.067) in control mice versus 0.108 (0.051–0.3550) in Alb-HMGB1−/−, p<0.05. Alb-HMGB1−/− tumors had significantly ameliorated proliferation rates and increased apoptosis as evident by immunofluorescent staining for Ki67 (median 112 (57–143) Ki67+ cells/106 μm2 in control mice versus 28 (4–47) Ki67+ cells/106 μm2 Alb-HMGB1−/− mice; p<0.01) and TUNEL (median 8.7 (2-12.3) TUNEL+ cells/103μm2 in control mice versus 15.4 (8–32.7) TUNEL+ cells/103 μm2 in Alb-HMGB1−/− mice; p<0.05), respectively. Nuclei (blue), Actin (white), PGC-1α/4HNE/Ki67 (red), TUNEL (green), Scale Bars 100μm. n=6 mice/group