Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2015 Apr 13;29(6):856–872. doi: 10.1210/me.2014-1349

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2015 by the Endocrine Society

PMC Copyright notice

Figure 1. — TH and CORT act synergistically to induce Klf9 mRNA in frog and mouse. A, Hormone-dependent Klf9 gene expression in X. laevis cell lines. We treated the X. laevis embryonic fibroblast (XTC-2) and renal epithelium (A6)-derived cell lines (n = 4–6/treatment) with vehicle, T₃ (5nM), CORT (100nM), or T₃ plus CORT for 4 hours before harvesting for RNA isolation. Hormone treatment caused statistically significant increases in Klf9 mRNA in both cell lines. XTC-2, T₃: 1.6-fold; CORT: 1.6-fold; T₃ + CORT: 5-fold; F_(3,15) = 85.01, P < .01; A6, T₃: 4-fold; CORT: 12-fold; T₃ + CORT: 53-fold; F_(3,15) = 93.59, P < .01; ANOVA. B, Hormone-dependent Klf9 gene expression in the mouse hippocampus-derived cell line HT-22. We treated HT-22 cells (n = 4–6/treatment) with vehicle, T₃ (30nM), CORT (100nM), or T₃ plus CORT for different times before harvesting for RNA extraction. Hormone treatment caused statistically significant increases in Klf9 mRNA (F_(3,15) = 42.36, P < .001; ANOVA): T₃: 2.4-fold; CORT: 2.7-fold; T₃ + CORT: 6.5-fold; 1 hour: F_(3,15) = 216.5, P < .001; 2 hours: F_(3,15) = 17.6, P < .01; 4 hours: F_(3,15) = 68.26, P < .001; 6 hours. C, Hormone-dependent Klf9 gene expression in early prometamorphic (Nieuwkoop-Faber stage 52–54) X. tropicalis tadpole brain (diencephalon). We treated tadpoles (n = 6/treatment) with T₃ and CORT at the doses indicated in the graph for 24 hours (hormones replenished at 12 hours), collected brains and microdissected the diencephalon for RNA extraction. Hormone treatment caused statistically significant increases in Klf9 mRNA (F_{(6, 41)} = 47.99, P < .001; ANOVA): 1nM T₃: 2.67-fold; 100nM CORT: 2-fold; 1nM T₃ + 100nM CORT: 5.75-fold; 5nM T₃ + 100nM CORT: 13.5-fold. D, Hormone-dependent Klf9 gene expression in the hippocampus of PND6 wild-type C57/BL6J mice. Mice were left unhandled, or were given ip injections of vehicle (corn oil + EtOH 0.05%), T₃ (25 μg/kg BW), CORT (14 mg/kg BW), or T₃ plus CORT. One hour after injection, mice were killed, and the hippocampal region was dissected for RNA extraction (n = 5–6/ treatment). Hormone treatment caused statistically significant increases in Klf9 mRNA (F_{(4, 25)} = 121.4, P < .01; ANOVA): T₃: 3-fold; CORT: 1.5-fold; T₃ plus CORT: 7-fold. In all experiments, total RNA was analyzed for Klf9 mRNA levels by RT-qPCR using TaqMan assays. The Klf9 mRNA levels were normalized to the reference genes rpL8 (frog) and Gapdh (mouse), which were unaffected by hormone treatment (data not shown). Tissue culture experiments were repeated 3–4 times. Bars represent the mean ± SEM, and letters above the means indicate significant differences among treatments (means with the same letter are not significantly different; Tukey's multiple comparison test; P < .05). Dashed lines indicate the calculated additive effect of combined hormone treatment.