Figure 1. Endogenous SOX9 is up-regulated in NT2/D1 cells in response to SRY.

(A) SOX9 is significantly upregulated in NT2/D1 cells transiently overexpressing SRY (1 µg), as measured by QRT-PCR and compared to empty vector (n = 3). (B) Immunostaining for endogenous SOX9 and exogenous SRY (Anti-Flag) reveals a positive correlation between SOX9 fluorescence and exogenous SRY fluorescence (R² = 0.835, n = 50). Scale bar represents 10 µm. Each point averages three fluorescence readings per NT2/D1 cell (Blue Diamond - SRY-transfected cells; Pink triangle - non-Flag cells). (C) (Upper panel) H3 Me₃K4 (red) is exclusively nuclear in NT2/D1 cells. Overlap of SRY (green) and H3 Me₃K4 in NT2/D1 cells reveals co-localization (yellow) within nuclear compartments. (Lower panel) SC-35 (red) is localized to alternate nuclear compartments, as overlap images of Flag-SRY and SC-35 staining do not show co-localization. DAPI (blue) stains nuclear DNA. Scale bar represents 10 µm. Error bars represent the standard error of mean values. Two-tail t-Test of paired sample means was performed. * P<0.05.