SEARCH

SEARCH BY CITATION

Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
HEP_24631_sm_suppinfofig1.tif5166KSupporting Information Figure 1. Loss of both HNF-6 and RBP-J is associated with visible hepatic necrosis on gross examination of liver. (A-B) Representative images of the left lobe of the liver at age P15 in control mice (A) without Alb-Cre transgene and DKO mice (B). Multiple foci of hepatic necrosis (B, arrow) are visible on gross examination of DKO mice liver. These foci appear bile stained. (A) Scale bar = 1mm.
HEP_24631_sm_suppinfofig2.tif3113KSupporting Information Figure 2. Formed IHBDs in DKO mice at P15 are located centrally within the hepatic lobe and stain positive for both DBA and CK19. Representative paraffin sections from the left lobe of P15 control (A) and DKO (B) mice stained for Dolichos biflorus agglutinin (DBA) and CK19 as a marker of BECs. In DKO mice (n=5), the only formed IHBDs were located centrally within the hepatic lobe and stained positive for both DBA and CK19. Peripheral CK19+/DBA- ducts and BECs were absent in DKO animals compared to control at age P15. (A) Scale bar = 10μm.
HEP_24631_sm_suppinfofig3.tif6745KSupporting Information Figure 3. BECs present in DKO mice at P3 and P15 demonstrate similar proliferation ratio to BECs in control mice of same age. (A) Representative paraffin sections from the left lobe in P3 and P15 control (without Alb-Cre transgene, of equal age) and DKO mice stained for Ki67 protein as a proliferation maker and CK19 as a BEC marker. (A) Arrow, dual labeled cell for Ki67 and CK19. (A) Scale bar, 10um. (B) The ratio of dual labeled Ki67/CK19+ to total CK19+ cells counted separated by age and genotype. There was no statistical difference in P3 Control versus DKO (P = 0.10) and P15 Control versus DKO (P = 0.15) proliferation ratio. For control P3, a minimum of 85 total CK19+ cells were counted from each control mouse (n=4), with a total of 1099 CK19+ cells counted for P3 control. For DKO P3, a minimum of 61 total CK19+ cells were counted from each DKO mouse (n=5) with a total of 763 CK19+ cells counted for P3 DKO. For control P15, a minimum of 260 total CK19+ cells were counted from each control mouse (n=3), with a total of 1085 CK19+ cells counted for P15 control. For DKO P15, a minimum of 110 total CK19+ cells were counted from each DKO mouse (n=3), with a total of 421 CK19+ cells counted for P15 DKO. (B) Statistical analysis was performed using a two-tailed Student's t-test comparing proliferative ratios of separate mice.
HEP_24631_sm_suppinfofig4.tif4213KSupporting Information Figure 4. Alb-Cre mediated loss of RBP-J is associated with an increase in HNF-6 protein expression. (A,B) Representative paraffin sections from the left lobe immunostained for HNF-6 from E16.5 control (A) without Alb-Cre transgene and E16.5 RBP KO (B). At E16.5, RBP KO mice demonstrate an increase in HNF-6 protein expression, corresponding to an increase in HNF-6 mRNA expression demonstrated by quantitative RT-PCR analysis of total liver RNA also at E16.5 (Fig. 1A). (A) Scale bar, 100μm.
HEP_24631_sm_suppinfotab1.doc37KSupporting Information Table 1. Real Time Quantitative RT-PCR Primers
HEP_24631_sm_suppinfotab2.doc34KSupporting Information Table 2. Primary and Secondary Antibodies

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.