Supplemental Data

Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Deletion of exon 1 of pik3CA in genomic DNA from POMC expressing regions of pik3CA POMCKO mice. (a) The 1500-bp region between primer binding sites becomes 947 bp in length when exon 1 is excised by cre recombinase. (b) Gels showing amplified genomic DNA from a pik3r1 POMCKO mouse. Hyp = DNA from whole hypothalamus, Pit = pituitary DNA with evidence of p110alpha deletion from POMC expressing corticotrophs, Cort = frontal cortex, Hipp = hippocampus, Olf = olfactory bulb DNA, Bs = Brainstem. Note that any POMC expression in brainstem was not high enough to elicit evidence of pik3CA exon 1 deletion. (b) Immunohistochemistry for β-endorphin was performed following ISHH for p85α or p110α in brain slices from targeted knockouts and littermate controls and the number of dual positive neurons were quantified throughout the rostral-caudal extent of the hypothalamus. Very stringent criteria were used to assess coexpression to minimize false positives; coexpression was considered present if silver grains clustered around the outline of the cell beneath. *P < 0.05, **P < 0.01