Volume 128, Issue 1, Pages 33-42 (January 2005) Relation between hepatocyte G1 arrest, impaired hepatic regeneration, and fibrosis in chronic hepatitis C virus infection  Aileen Marshall, Simon Rushbrook, Susan E. Davies, Lesley S. Morris, Ian S. Scott, Sarah L. Vowler, Nicholas Coleman, Graeme Alexander  Gastroenterology  Volume 128, Issue 1, Pages 33-42 (January 2005) DOI: 10.1053/j.gastro.2004.09.076 Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 1 Immunofluorescent double labeling for cell cycle phase markers. (A) Cyclin D1 (green) and cyclin A (red) in hepatocellular carcinoma; colocalization not seen. (B) Cyclin A (green) and cytoplasmic cyclin B1 (red) in regenerating liver; colocalization seen in <2% of cyclin A-positive hepatocytes. (C) Cytoplasmic cyclin B1 (green) and PH3 (red) in hepatocellular carcinoma; colocalization only when nuclear cyclin B1 was also present. (D) Cyclin A (green) and PH3 (red) in regenerating liver; colocalization in <5% of cyclin A-positive cells. Arrow indicates cell positive for both cyclin A and PH3. Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 2 Immunoperoxidase stain in a liver biopsy specimen from a patient with chronic hepatitis C infection using cell cycle phase-specific antibodies. (A) Mcm-2, present throughout cell cycle. (B) Cyclin D1, maximal during G1. (C) Cyclin A, maximal during S phase. (D) Cyclin B1, cytoplasmic during G2. (E) PH3, detectable during mitosis. Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 3 Immunoperoxidase stain in a liver biopsy specimen from a patient with liver regeneration following transplant-related acute ischemic-reperfusion injury using cell cycle phase-specific antibodies. (A) Mcm-2, present throughout cell cycle. (B) Cyclin D1, maximal during G1. (C) Cyclin A, maximal during S phase. (D) Cyclin B1, cytoplasmic during G2 (closed arrow). Cyclin B1 translocates to the nucleus at the start of mitosis (open arrow). (E) PH3, detectable during mitosis. Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 4 Hepatocyte Mcm-2 and cell cycle phase markers comparing chronic HCV and liver regeneration. (A) Percentage Mcm-2-positive hepatocytes in HCV versus regeneration (P = .002). B–E give the median of results expressed as percentage of Mcm-2-positive hepatocytes for each case, termed labeling fraction. (B) Cyclin D1 (P = .2). (C) Cyclin A (P < .0001). (D) Cyclin B1 (P < .0001). (E) PH3 (P < .0001). For these and all subsequent figures, the black bar in the middle of the box represents the median, the box stretches between the lower and upper quartile, and the whiskers extend to the range of the data or 1.5 times the box length, whichever is shorter. Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 5 Hepatocyte Mcm-2 expression in chronic HCV liver biopsy specimens in relation to fibrosis stage according to Ishak’s criteria (P = .001; Jonckheere-Terpestra test). Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 6 Hepatocyte Mcm-2 expression comparing HCV antibody-positive, HCV RNA-negative individuals with HCV antibody positive, HCV RNA-positive patients (P = .001; Mann-Whitney U test). Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 7 Hepatocyte p21 expression in chronic HCV liver biopsy specimens according to fibrosis stage (P < .0001; Jonckheere-Terpestra test). Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions

Figure 8 Hepatocyte p21 expression comparing HCV antibody-positive, HCV RNA-negative individuals with HCV antibody- positive, HCV RNA-positive patients (P = .004; Mann-Whitney U test). Gastroenterology 2005 128, 33-42DOI: (10.1053/j.gastro.2004.09.076) Copyright © 2005 American Gastroenterological Association Terms and Conditions