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. 2022 Dec 2;13:7448. doi: 10.1038/s41467-022-34991-2

Fig. 3. NKX2-3 regulates St6gal1 expression via a conserved NCCE.

Fig. 3

A Schematic of the St6gal1 gene and an NCCE in the second intron. B EMSA showing migration of NKX2-3, COUP-TFII-GST (red arrows), and NKX2-3:COUP-TFII heterodimer (blue arrow) bound to an St6gal1 CE probe. C St6gal1 expression in DN-MAML bEnd.3 cells stably transfected with Nkx2-3 or Nr2f2 shRNA, evaluated by real-time PCR. D St6gal1 expression in DN-MAML bEnd.3 cells stably transfected with NKX2-3 or the NKX2-3 TN domain. E Activity of luciferase reporter driven by St6gal1 NCCE-containing promoter (St6Gal1-LUC; gray bars) or St6gal1 NCCE-containing promoter with mutated COUP-TFII binding sequence (St6Gal1_ΔCOUP-LUC; white bars), when co-transfected with Nr2f2 and/or Nkx2-3 in 293T cells. F Immunofluorescence of alpha-2,6-sialic acid binding lectin SNA staining in PP HEV in WT vs Nkx2-3 deficient mice (red). HEV are marked by AF450-labeled anti-addressin antibodies MECA79, MECA89, and MECA367. Autofluorescence is shown in green. Scale bars: 50 µm. Results in C-E are shown as mean ± SEM from three independent transfections. Two tailed t-test, paired. *: p value<0.05; **: p value<0.01; ***: p value < 0.005.