Fig. 8: LINC00629 activates NUCKS1/ASNS axis via sponging miR-4768-3p.

A The potential miRNAs targeted NUCKS1 were predicted from miRwalk database (black line) and Targetscan database (blue line). The potential miRNAs targeted NUCKS1 were predicted from miRDB (red line). The five overlapped miRNAs were obtained. B The binding site of the five miRNAs on LINC00629 were listed and the effects of miRNAs on LINC00629 were assessed. C The indicated miRNAs were transfected into MNNG/HOS and 143B cells. The expression of NUCKS1 was detected by western blot. D miR-4768-3p inhibitor were transfected into MNNG/HOS and 143B cells. The expression of NUCKS1 was detected by western blot. E Potential binding site of miR-4768-3p on NUCKS1 3’UTR was shown. The wild-type binding site or mutant of miR-4768-3p on NUCKS1 3’UTR were cloned into a pSICHECK2 vector. The blue indicates the mutated region. F The WT or Mut of NUCKS1 3′UTR was respectively transfected into MNNG/HOS cells with or without miR-4768-3p overexpression. The luciferase activity of NUCKS1 3′UTR was measured. G The WT and Mut of NUCKS1 3’UTR was transfected into MNNG/HOS cells with or without miR-4768-3p inhibitor treatment. The luciferase activity of NUCKS1 3’UTR was measured. H Schematic illustration of pSICHECK2 reporter constructs used in luciferase assays to examine the effects of miR-4768-3p on luciferase activity of LINC00629 and different truncations named BS1 and BS2. I the full length of LINC00629 and the different truncations were transfected into MNNG/HOS cells with or without miR-4768-3p overexpression. The luciferase activity was measured. J The BS2 wild type and mutant were constructed into pSICHEK2 vector. K, L BS2 WT and Mut were transfected into MNNG/HOS cells with or without miR-4768-3p or miR-4768-3p inhibitor. The luciferase activity was measured. Data in B, F, G, I, K and L were analyzed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001.