Figure 5

PITX2A, PITX2B1 and PITX2B2 isoforms trans -activate gonadotrope-specific genes. A, C and D) CV-1 cells seeded in 12-well plates were co-transfected with the indicated promoter-reporters (0.9 μg/well) along with empty (pcDNA3.0) or PITX2 expression vectors (0.3 μg/well). B) The Lhb promoter-reporter (0.9 μg/well) was co-transfected with PITX2 expression vectors plus or minus SF-1 (both expression vectors at 0.3 μg/well; DNA in all wells was balanced with pcDNA3.0). After approximately 30 h, lysates were collected for luciferase assays. Data were derived from three to five experiments performed in triplicate. Bars with different symbols differ significantly. E) Immunoblot showing relative PITX2A, B1 and B2 expression in CV-1 cells, whole cell extracts run on a 4–12% Nupage^® Novex Bis-Tris gel with MOPS buffer.