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. 2021 Jan 15;10:581459. doi: 10.3389/fonc.2020.581459

Figure 2.

Figure 2

HEYL controls the expression of multiple angiogenic factors. (A) Western blot analysis of Myc-tagged HEYL expression in HS578T-tet-off-HEYL at 0, 6, and 24 h after doxycyline (dox) withdrawal. At the 24 h time point, Dox was re-added, and Myc-tagged HEYL expression was examined 48 h later (72 h after initiation of the experiment). (B) A scatter plot (showing log2-fold change) and heatmap show genes with expression changes at 6 and 24 h after HEYL induction relative to 0 h cells (uninduced cells) (also listed in Supplementary Table 1 ). (C) RT-qPCR validation of expression of major cytokines in the HS578T-tet-off-HEYL-inducible cells in (B) at 0, 6, 24, and at the 72 h time point (48h after re-induction of HEYL expression). mRNA expression levels relative to 36B4, a ribosomal gene, are shown. (D) Antibody array-based detection of cytokines in conditioned medium from HS578T-tet-off-HEYL-induced and –uninduced cells (6h); +C are quality controls (E) Western blot analysis of HEYL in MDA-MB-231-shRNA cells and re-expression after introduction of shRNA-resistant, Myc-tagged HEYL. (F) qPCR of cytokine expression in the MDA-MB-231 cell panel in (E). mRNA expression levels relative to 36B4, a ribosomal gene, are shown. *P < 0.05; **< 0.05; ***< 0.001.