Figure 4

miR-135a regulation of HOXA10 expression. (A) Images of GFP-labeled HEK293 cells (left panel). Western blot of GFP protein in GFP-labeled HEK293 cells (right panel). Anti-GAPDH antibody was used as a loading control (bottom panel). The HEK293 cells were co-transfected with plasmids pEGFP-HOXA10 and pSilencer-miR-135a (pS-135a) or pEGFP-HOXA10 and pSilencer4.1CMV-negative (pS-N) respectively. (B) Western blot of HOXA10 protein in HEK293 cells transfected with pS-135a or pS-N. Anti-actin antibody was used as a loading control (bottom panel). (C) HOXA10 mRNA expression in pS-135a or negative transfected HEK293 cells detected by SYBR Green qRT-PCR. β-actin was used as a loading control. (D) Western blot of HOXA10 protein in MCF-7 cells transfected with pS-135a or pS-N. Anti-GAPDH antibody was used as a loading control (bottom panel). (E) Western blot of HOXA10 protein in BT549 cells transfected with inhibitor control (inhibitor NC) or 135a inhibitor, pS-135a or pS-N. Anti-GAPDH antibody was used as a loading control (bottom panel). n = 3, *, p < 0.05, pS-135a significantly decreased mRNA levels (normalized to β-actin mRNA) compared with cells transfected with pS-N.