Fig. S1.

Tet1 expression negatively correlates with gonadotrope differentiation (Fig. 1). (A and B) qPCR analyses of relative levels of each of the TET enzyme mRNAs in (A) primary gonadotropes from immature (6 d) or mature (8–14 wk) mice or (B) partially differentiated (αT3-1) or fully differentiated (LβT2) gonadotrope cell lines; Tet mRNA levels were calculated by using standard curves of plasmid cDNA (n = 3–9). A t test was used to compare levels of the same gene between groups of mice (**P < 0.01 and ***P < 0.001; NS indicates P > 0.05). (C) TET1 (red) and CGA (green) in the pituitary of 5-d-old female mouse shown at lower magnification (Top Left) and subsequently the boxed region enlarged for each color channel separately or overlaid (Bottom Right). (Scale bars: lower magnification, 100 µm; higher magnification, 50 µm.) Notably, TET1 is seen exclusively in CGA-expressing cells at this stage, but is not found in all CGA⁺ cells. (D and E) Tet2, Tet3, and Fshb mRNA levels in the OVX (D) or castrated mice (E) in Fig. 1 G and H shown relative to levels in the intact mice; there are no significant differences between means of the two groups for any of these genes (P > 0.05). (F) Gonadotrope cells from intact females (freely cycling) and OVX mice were cultured with E₂ for 48 h before qPCR analysis of Tet1 mRNA levels as before (n = 3; *P < 0.05; NS indicates P > 0.05).