Fig. 2 | Oncogene

Fig. 2

From: Estradiol induces BDNF/TrkB signaling in triple-negative breast cancer to promote brain metastases

Fig. 2

E2 upregulates BDNF in reactive astrocytes in vitro and in vivo. a Human primary astrocytes were treated with vehicle ethanol (OH) (Hu-Ast-OH) or 10 nM E2 (Hu-Ast-E2) for 24 h and cell lysates were assessed using growth factor arrays. Graph shows mean relative intensity of growth factors and cytokines with >1.4-fold increase in E2 compared to OH-treated astrocytes (n = 2 samples in independent arrays). b Primary mouse astrocytes were treated with vehicle (OH) or 10 nM E2 for the indicated times and BDNF mRNA levels were measured using qRT-PCR. Dots represent fold change mRNA levels normalized to β-Actin mRNA and relative to OH-treated. Line shows mean. The data were analyzed using Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Adjusted P values are shown (n = 4). c WB shows pro-BDNF (34 kD) levels in mouse astrocytes (mAst) treated with vehicle (OH), 10 nM E2 alone or in combination with 1 µM Tam for 72 h. Numbers indicate ratio of BDNF/α-tubulin relative to OH-control (n = 2). d Double IF staining of BDNF (green) and reactive astrocytes (GFAP, red) in brain sections of OVX or E2-treated mice injected with 4T1BR5 cells. Left: representative image of GFAP+/BDNF+ cells in brains of OVX and E2-treated mice. The box indicates a region of interest shown at 4X magnification. Scale bar is 100 µm. Right: Dots show the mean percentage of GFAP+ cells expressing BDNF in 9 × 0.134 mm2 fields per mouse in OVX (n = 10) and E2-treated mice (n = 12). Two-tailed Mann–Whitney test was used for analysis. Line shows the group mean

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