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dvdy21437-DVDY21437Anderson_et_al._FigureS1.tif1097K Supplemental Fig. 1.Expression ofCldn6in adult tissues. RT-PCR on various adult tissues show expression ofCldn6in 11 of the 23 tissues examined after 35 rounds of amplification. E7.5 embryo cDNA was used as a positive control.β-actinwas used as a loading control.
dvdy21437-DVDY21437Anderson_et_al._FigureS2.tif1150K Supplemental Fig. 2.The CIHV cassette driven by the pCAGGS promoter expresses nuclear localized Venus and has tamoxifen-dependent Cre recombinase activity. pCAGGS-CIHV was transfected into COS cells. Transfected cells expressed H2B/Venus. The pCAGGS-CIHV plasmid was also co-transfected with a pCAGGS-floxed stop- RFPplasmid (shown schematically; triangles represent loxP sites). RFP expression was only seen in cells grown in the presence of tamoxifen (+ tamoxifen). Exposure in the RFP channel was equivalent for both images. TM, tamoxifen.
dvdy21437-DVDY21437Anderson_et_al._FigureS3.tif562K Supplemental Fig. 3.Targeting strategy for theCldn6locus. TheCldn6gene is comprised of three exons, with exon 3 containing the entire 660-bp coding sequence. The coding sequence was removed and replaced at the endogenous ATG translational start site with the CIHV cassette (see text and Experimental Procedures section for details). As a result, no intronic sequences that may harbor regulatory regions were deleted. Color scheme: orange bar,Cre-ER T2 ; magenta bar, IRES; green bar,H2B/Venus ; red oval, SV40 polyadenylation signal; blue bar,ACNcassette; blue triangles, loxP sites.
dvdy21437-DVDY21437Anderson_et_al._FigureS4.tif615K Supplemental Fig. 4.Strategy for lineage analysis.A:Schematic of the cross betweenCldn6 CIHV/+ and R26R lines.B:Table of tamoxifen administration with probable corresponding time of Cre-mediated recombination shown as a blue line (based on Hayashi and McMahon, 2002). TM, tamoxifen; βgeo, lacZ/neomycin resistance fusion gene.

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