Fig 1: ZEB2 is a target gene of miR-132. (A) HN1 was predicted as a target gene of miR-132 with a potential miR-132 binding site located in the HN1 3'-UTR. (B) A549 and (C) H460 cells transfected with miR-132 mimic and the WT or MUT 3'-UTR of HN1 exhibited no change in their luciferase activity. (D) ZEB2 was predicted as a target gene of miR-132 with a potential miR-132 binding site located in the ZEB2 3'-UTR. miR-132 mimic decreased the luciferase activity of the WT but not the MUT ZEB2 3'-UTR in (E) A549 and (F) H460 cells. *P<0.05 vs. NC group. ZEB2, zinc finger E-box binding homeobox 2; miR-132, microRNA-132; HN1, neurological expressed 1; 3'-UTR, 3'-untranslated region; WT, wild-type; MUT, mutant; NC, negative control.
Fig 2: Effect of miR-132 and hesperidin on ZEB2 expression in A549 and H460 cells. (A) In A549 cells, miR-132 mimic demonstrated no effect on the mRNA expression of HN1, whereas HN1 siRNA reduced the mRNA expression of HN1. (B) In A549 cells, miR-132 mimic and ZEB2 siRNA both decreased the mRNA expression of ZEB2. (C and D) In A549 cells, miR-132 mimic demonstrated no effect on the protein expression of HN1, but reduced the protein expression of ZEB2. (E) In A549 cells, miR-132 mimic and ZEB2 siRNA both decreased the protein expression of ZEB2. (F) Treatment with hesperidin exhibited no effect on the mRNA expression of HN1 in A549 cells. (G) Treatment with hesperidin decreased the mRNA expression of ZEB2 in a concentration-dependent manner in A549 cells. (H) Western blot results of HN1 and ZEB2 protein in A549 cells treated with different concentrations of hesperidin. (I) Treatment with hesperidin demonstrated no effect on the protein expression of HN1 in A549 cells. (J) Treatment with hesperidin decreased the protein expression of ZEB2 in a concentration-dependent manner in A549 cells. (K) In H460 cells, miR-132 mimic had no effect on the mRNA expression of HN1, whereas HN1 siRNA reduced the mRNA expression of HN1. (L) In H460 cells, miR-132 mimic and ZEB2 siRNA both decreased the mRNA expression of ZEB2. (M and N) In H460 cells, miR-132 mimic exhibited no effect on the protein expression of HN1, but HN1 siRNA reduced the expression. (O) In H460 cells, miR-132 mimic and ZEB2 siRNA both decreased the protein expression of ZEB2. (P) Treatment with hesperidin demonstrated no effect on the mRNA expression of HN1 in H460 cells. (Q) Treatment with hesperidin decreased the mRNA expression of ZEB2 in a concentration-dependent manner in H460 cells. (R) Western blot results of HN1 and ZEB2 protein in H460 cells treated with different concentrations of hesperidin. (S) Treatment with hesperidin had no effect on the protein expression of HN1, (T) but decreased the protein expression of ZEB2 in a concentration-dependent manner in H460 cells. *P<0.05 vs. control/untreated group. ZEB2, zinc finger E-box binding homeobox 2; miR-132, microRNA-132; HN1, neurological expressed 1; siRNA, small interfering RNA.
Fig 3: Effect of hesperidin on the tumor volume and colony formation efficiency of NSCLC, and on HN1 and ZEB2 expression in rats transplanted with NSCLC cells. (A) The tumor volume in rats transplanted with NSCLC cells and treated with hesperidin was smaller compared with that in rats transplanted with NSCLC cells alone. (B) Treatment with hesperidin significantly reduced the colony formation efficiency of NSCLC cells. (C) Hesperidin increased miR-132 expression in the rats. (D) Hesperidin decreased the mRNA expression of ZEB2 in rats transplanted with NSCLC cells. (E) Hesperidin demonstrated no effect on the mRNA expression of HN1 in rats transplanted with NSCLC cells. (F) Hesperidin decreased the protein expression of ZEB2 in rats transplanted with NSCLC cells, but showed no effect on the protein expression of HN1. (G) Quantification of the western blots demonstrated that hesperidin significantly decreased the protein expression of ZEB2 in rats transplanted with NSCLC cells. (H) Quantification of the western blots demonstrated hesperidin had no effect on the protein expression of HN1 in rats transplanted with NSCLC cells. (I) Hesperidin increased the apoptosis of NSCLC cells (magnification ×200). *P<0.05 vs. NC group. NSCLC, non-small cell lung cancer; ZEB2, zinc finger E-box binding homeobox 2; miR-132, microRNA-132; HN1, neurological expressed 1; NC, negative control.
Supplier Page from Abcam for Anti-HN1 antibody [EPR7364]