Fig. S2

Aldh1-expressing cells and Notch-responsive cells represent distinct cell types in adult zebrafish pancreas. (A-D) FACS sorting was performed on single cells isolated from adult pancreas of wild type (A and B) or Tp1:eGFP (C and D) fish. Wild-type cells were labeled with Aldefluor and then FACS sorted, while cells from Tp1:eGFP fish were sorted for eGFP. Sorted populations were then subjected to cytospin and immunofluorescent labeling for Aldh1. In the case of cells from Tp1:eGFP fish, labeling was also performed for eGFP. Note that Aldh1 protein is detected in Aldefluor^pos but not Tp1:eGFP^pos cells, while eGFP is detected in Tp1:eGFP^pos but not Aldefluor^pos cells. Arrows in (C) indicate low-abundance Aldh1-sorted cells present in Tp1:eGFP^neg but not Tp1:eGFP^pos cell fraction. E, RT-PCR analysis of gene expression in FACS sorting populations. Positive and negative populations of cells sorted for either Aldh1 activity or Tp1:eGFP expression displayed differential expression of aldh1a2, prom1, and sca1, with contrasting patterns of enrichment in Aldefluor^pos vs. Aldefluor^neg and Tp1:eGFP^pos vs. Tp1:eGFP^neg cell fractions, further documenting the non-overlapping nature of Aldh1-expressing and Notch-responsive progenitor cells. Note that both populations are depleted of transcripts encoding differentiated endocrine (insulin), acinar (elastase1) and ductal (krt18) markers.