Supplementary Figure 12: Generation of human iPS cell lines harboring copGFP and BCL2 knock in insertions.
From: Generation of human endothelium in pig embryos deficient in ETV2
(a) Schematic outlining the knock in strategies for copGFP to the CLYBL locus (top) and BCL2 over expression to the AAVS1 locus (bottom) (CF1/CR1;CF2/CR2). (b) PCR gel images showing correct targeting of RhiPSC11 cGFP in the CLYBL locus (5’CF1/CR1 & 3’CF2/CR2 CLYBL locus targeting validation) and RhiPSC11 cGFP-BCL2 both in the CLYBL and AAVS1 loci (5’AF1/AR1 & 3’AF2/AR2 AAVS1 locus targeting validation), yet not in RhiPSC11 WT nor NTC (no template control). Data shown are representative of two independent PCR verifications of the correctly targeted lines. (c) Immunoblot image showing over expression of BCL2 in RhiPSC11 cGFP-BCL2 and the positive control (PC) but not in the WT hiPSCs. Data shown are representative of two independent immunoblot verifications of the correctly targeted lines.
