The document summarizes immunofluorescence staining results for CFDA and CK19 in liver samples from various mouse models and timepoints. In DKO P58 mice, hilar ducts typically co-stained for CFDA and CK19 while peripheral areas showed disorganized CK19-positive cells lacking CFDA. A sicker DKO P58 mouse had fewer disorganized CK19 cells and some hilar ducts with weak CFDA staining. A P70 HNF-6 KO mouse showed normal duct staining. P70 NICD/HNF-6 mice had well-formed ducts positive for CFDA but multiple peripheral and hilar areas lacking CFDA, associated with disorganized CK19-

1. CFDA/CK19 ImmunoflourescenceP58 DKO n=2P70 HNF-6 KO n=1P70 NICD/HNF-6 n=3

2. DKO P58: 1639 P58 male, T bili 7.9, weight 25.80gmsCFDACK19Hilar ducts typically with good CFDA co-stainMultiple areas in periphery with disorganized CK19 positive cells not CFDA positivePictures intralobular to periphery

3. DKO P58: 1540 P58 male, T bili39.7, weight 13.11gms, very ill appearingCFDACK19Much fewer areas of disorganized CK19 cellsSome hilar ducts with poor CFDA stainOverall, very limited CFDAHilar duct negative with small duct adjacent postive for CFDA

4. P70 HNF-6 KOCFDACK19NormalAll ducts with CFDANo ducts without CFDA

5. P70 NICD/HNF-6: 1575CFDACK19Most well formed ducts positive for CFDAMultiple areas along periphery and hilum without CFDA positive staining, mostly if associated with areas of disorganized CK19+ cells

6. P70 NICD/HNF-6: 1876CFDACK19Most well formed ducts positive for CFDAMultiple areas along periphery and hilum without CFDA positive staining, mostly if associated with areas of disorganized CK19+ cells