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. 2019 Apr 1;6(2):ENEURO.0400-18.2019. doi: 10.1523/ENEURO.0400-18.2019

Figure 2.

Figure 2.

Vglut2-ires-Cre-specific recombination of the Pomc FNΔ2 allele. A, Schematic representing the WT Pomc locus (control), the presence of the Cre-excisable floxed neomycin cassette along with the knock-out of nPE2 (FNΔ2), and the FNΔ2 allele after Cre-mediated excision leaving only the knock-out of nPE2 (restored). B, Hybridization location of the three oligonucleotide primer set used to assess the integrity of the Cre-mediated recombination. The forward 1 (F1) primer hybridizes upstream of nPE2, the forward 2 (F2) primer hybridizes to the intact neomycin cassette, and the reverse (R) primer is specific to the knocked out nPE2 locus. C, Verification of Cre-mediated genetic excision of the floxed-neomycin cassette from the Pomc neural enhancer locus in restored mice. The intensity of the recombined band (PCR product of F1 and R; 287 bp) versus the non-recombined band (PCR product of F2 and R; 180 bp) was much stronger in the medial basal hypothalamus than in the dorsal striatum. (P < 0.00001, see Table 2).