Figure 1.
Clustergram showing the expression profile of significantly (p<0.05) altered genes related to the cell cycle in adrenal ZF fraction or ZG fraction after inhibition of the HPA axis with DEX for two days (50 µg/100 g BW) (A) Correlation of up- or down-regulated genes in ZF fraction and ZG fraction; genes regulated in both DEX and control treatments are presented in the figure intersection (B).
Analysis of Nek2b protein expression in adrenal ZF fraction and ZG fraction after inhibition of HPA axis with DEX (C). Adrenal glands were separated in two fractions (ZG and ZF), and total protein or total RNA, respectively, was extracted for the immunoblotting or PCR array analysis. Control animals received saline only (Control). *p<0.05, n = 3.
Figure 2.
Immunolocalization of Nek2b expression in the adrenal cortex after HPA inhibition.
Adrenal sections were obtained from rats treated with: (A) saline only or (B) DEX for two days (50 µg/100 g BW). Nek2b-positive cells were stained brown. Sections were counterstained with Harris’ hematoxylin and differentiated with a saturated solution of lithium carbonate. Bars 10 µm.
Table 1.
Genes significantly regulated in the ZG and ZF fractions of the adrenal cortex after inhibition of the HPA axis.
Figure 3.
Results of the PCR array for cell cycle related genes in the ZG fraction.
Clustergram showing the expression profile of significantly (p<0.05) altered genes related to the cell cycle in the adrenal ZG fraction after inhibition of HPA axis with DEX for two days (50 µg/100 g BW), followed by ACTH or N-POMCMet or N-POMCCys treatments (A). Correlation of up- or down-regulated genes in the ZG fraction after treatments, showing the number of genes regulated by both treatments with POMC-derived peptides (B). Up- or down-regulated genes related to specific cell cycle phases after ACTH, N-POMCMet, or N-POMCCys treatments (C). G1: G1 phase and G1/S transition; G2: G2 phase; S: S phase and DNA replication; M: M phase.
Figure 4.
Results of the PCR array for cell cycle related genes in the ZF fraction.
Clustergram showing the expression profile of significantly (p<0.05) altered genes related to the cell cycle in the adrenal ZF fraction after inhibition of the HPA axis with DEX for two days (50 µg/100 g BW), followed by ACTH, N-POMCMet, or N-POMCCys treatments (A). Correlation of up- or down-regulated genes in the ZF fraction after treatments, showing the number of genes regulated by both treatments with POMC-derived peptides (B). Up- or down-regulated genes related to specific cell cycle phases after ACTH, N-POMCMet, or N-POMCCys treatments (C). G1: G1 phase and G1/S transition; G2: G2 phase; S: S phase and DNA replication; M: M phase.
Table 2.
Genes significantly regulated in the ZG fraction of the adrenal cortex after inhibition of the HPA axis followed by treatments with ACTH, N-POMCMet, or N-POMCCys.
Table 3.
Genes significantly regulated in the ZF fraction of the adrenal cortex after inhibition of the HPA axis followed by treatments with ACTH, N-POMCMet, or N-POMCCys.
Figure 5.
Analysis of Nek2b protein expression in adrenal ZF fraction and ZG fraction after inhibition of the HPA axis with DEX, followed by ACTH, N-POMCMet, or N-POMCCys treatments.
Adrenal glands were separated in two fractions (ZG and ZF), and total protein was extracted for immunoblotting analysis (A). Immunolocalization of Nek2b expression in the adrenal cortex after HPA inhibition followed by ACTH (B), N-POMCMet (C), or N-POMCCys (D) treatments. Adrenal sections were obtained from rats treated previously with DEX for two days (50 µg/100 g BW). Nek2b-positive cells were stained brown. Sections were counterstained with Harris’ hematoxylin and differentiated with a saturated solution of lithium carbonate. Bars 10 µm. *p<0.05, n = 3.
Figure 6.
Immunolocalization of Notch2 expression in the adrenal cortex.
Adrenal sections were obtained from rats treated with saline only. Notch2-positive cells are stained brown and indicated by arrows (A). Negative control without primary antibody (B). Sections were counterstained with Harris’ hematoxylin and differentiated with a saturated solution of lithium carbonate. Bars 10 µm.
Figure 7.
Immunolocalization of Notch1 expression in the adrenal cortex.
Adrenal sections were obtained from rats treated with saline only (A and B) or two days with DEX (50 µg/100 g BW) followed by: saline (C), or ACTH (D), or N-POMCMet (E), or even N-POMCCys (F). Notch1-positive cells are stained in brown and indicated by arrows. Sections were counterstained with Harris’ hematoxylin and differentiated with a saturated solution of lithium carbonate. Bars 10 µm.
Figure 8.
Immunolocalization of Notch3 expression in the adrenal cortex.
Adrenal sections were obtained from rats treated with saline only (A and B) or with DEX for two days (50 µg/100 g BW) followed by: saline (C), or ACTH (D), or N-POMCMet (E), or even N-POMCCys (F). Notch3-positive cells are stained in brown and indicated by arrows. Sections were counterstained with Harris’ hematoxylin and differentiated with a saturated solution of lithium carbonate. Bars 10 µm.