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. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: J Mol Endocrinol. 2015 Nov 12;56(2):55–67. doi: 10.1530/JME-15-0171

Figure 2. Epo treatment induces proliferation, POMC expression, and JAK2- and STAT3-signaling in WT hypothalamus NPC-culture.

Figure 2

(A-B) Dose-dependent increase in POMC mRNA levels relative to untreated cells and adjusted to β-Actin mRNA levels (A), and POMC protein normalized to β-Actin after 24 h of Epo-treatment (B) were determined. Densitometry data is representative of three independent experiments. (C) Western blotting for phosphorylated-JAK2 (p-JAK2), total JAK2 (t-JAK2), phosphorylated-STAT3 (p-STAT3), total STAT3 (t-STAT3), and β-Actin protein in control- and Epo-treated (20 U/ml) NPC cultures, and quantitative densitometry analysis of band intensity ratio of p-STAT3 and t-STAT3 are shown. Densitometry data is representative of three independent experiments. (D-E) Level of POMC (D) and β-tubulin (E) mRNA as measured by real time RT-PCR analysis of NPCs after saline-treatment (Cont), Epo-treatment, Epo with STAT3-inhibitor WP1066, and WP1066 only. The mRNA expression levels are relative to saline–treated (Cont.) cells and adjusted to β-Actin mRNA levels, and represents mean ± SD from triplicate reactions. (F) Proliferation of NPC cultures after 32 h and 48 h of Epo-treatment at 0, 10, and 20 U/ml doses. Relative cell numbers are representative of triplicate treatments for each dose. Statistical significance is indicated by *p<0.05, and **p<0.01, (one-way ANOVA followed by Bonferroni correction).