Skip to main content
. 2019 Apr 6;9(7):2084–2099. doi: 10.7150/thno.29463

Figure 6.

Figure 6

HOXB13 targets to the promoter of Slug and upregulates Slug gene expression. (A) H1299 and A549 lung adenocarcinoma cells were transiently transfected by Flag-HOXB13 or GFP-HOXB13 separately, controlled by Flag or GFP. Cell lysates were prepared and were subjected to Western blot analyses using the anti-Slug antibody. (B) HOXB13 occupies Slug promoter. Potential regions of the Slug promoter with HOXB13 binding sites (double arrow). ChIP analysis was performed using either an anti-HOXB13 ChIP-grade antibody or control IgG in H1299 Flag-HOXB13 cells. Gel picture of ChIP analysis for HOXB13 occupancy on Slug promoter was given. Sites 1 to 5 are enriched on Slug promoter under HOXB13 expression determined by qPCR analysis. (C) Slug is required for HOXB13-promoted lung adenocarcinoma cell migration. Upper panel: H1299 cells were transfected by Flag-HOXB13 controlled by Flag vector, simultaneously cells were transfected with Slug siRNA controlled by scramble RNA. Cell migration was determined and results were analyzed from three independent experiments (Unpaired Student's t-test, *p<0.05, **p<0.01). Lower panel: Western blot analysis was subjected to determine the upregulation of Slug by HOXB13 overexpression and Slug knockdown by siRNA using an anti-Slug antibody.