The regulatory effect of HOXD8 and Lhx3 on RP11-269F21.2, DIAPH2-AS1, and RP11-445K13.2
(A) HOXD8 and Lhx3 were knocked down in HTR-8/SVneo cells under hypoxia using RNA interference technology. (B) The expression of RP11-269F21.2, DIAPH2-AS1, and RP11-445K13.2 expression in HTR-8/SVneo cells under hypoxia was measured by PCR assay after HOXD8 and Lhx3 knockdown. (C) A wild-type sequence (-100 ∼ -900 bp) containing all putative binding sites of HOXD8 on the promoter of DIAPH2-AS1 gene was cloned into pGL3 luciferase report. Two of the binding sites were mutated to established two mutant vectors. The vectors were transfected into HTR8/SVneo cells alone or with HOXD8 over-expression vector. *P<0.05 and **P<0.01. WT: wild-type vector; MT: mutant-type vector; Ove: HOXD8 overexpression vector. KD: knockdown