Fig 1. Generation of germline HA-hPIT1^tg/+ transgenic mouse.

Scheme for recombination of the targeting vector pBT478.6-mKozak-CAG-LSL-HA-hPIT1 TARGATT into the H11 locus (A). Primer positions are shown, which were used in (B) to detect the modified H11attR site at 5’ junction (transgene inserted between attP1 and attP2) and 3’ junction (transgene inserted between attP2 and attP3) using primers from the Applied StemCell kit (primers 1,3 and 2,4, respectively). Germline excision shown in (C) was confirmed in (D) using primers detecting the wildtype H11 site (primers 896 and 897), the inactive LSL-positive (primers 879, 880) and the active germline ΔLSL alleles (primers 883 and 880). Primers 823 and 824 detect Cre recombinase, which is absent in the two mice shown consistent with germline excision of the LSL cassette of the mouse sample run in the left lane.