Fig. 4

Linc01296 directly bound miR-26a and regulated GALNT3 expression. a MiR-26a expression was detected in CRC tumor tissues and corresponding nontumor tissues. b MiR-26a showed a differential level in CRC cells. c A negative correlation between miR-26a and linc01296 was determined by Spearman’s correlation analysis. d Sequence aligment of miR-26a with putative binding sites in the wild-type and mutant-type regions of linc01296 (left panel) was shown. The wild-type and mutant miRNA target linc01296 sequences were fused with luciferase reporter, transfected with miRNA mimic and NC mimic (right panel). e qRT-PCR and IHC analysis were carried out to detect GALNT3 expression. f Differential GALNT3 level was found by qRT-PCR and western blot in CRC cells. g Immunofluorescence staining of GALNT3 was detected. Red fluorescence: GALNT3, blue fluorescence: DAPI. h Relationship between miR-26a and GALNT3 mRNA expression was analyzed by Spearman’s correlation in CRC tissues and the corresponding normal tissues. i Sequence aligment of miR-26a with predicted binding sites in the wild-type and mutant-type regions of GALNT3 (left panel) was shown. The wild-type and mutant type GALNT3 sequences were fused with luciferase reporter, transfected with miRNA mimic and NC mimic (right panel). j The co-precipitated RNA was detected by qRT-PCR in RNA immunoprecipitation experiment. Linc01296 and miR-26a were presented as fold enrichment in Ago2 relative to IgG immunoprecipitate. k The effort of linc01296 in CRC cells was detected by qRT-PCR and western blot. l Silinc01296 regulating GALNT3 expression of CRC cells was measured. m GALNT3 expression was detected in CRC cells transfected with linc01296 and miR-26a mimic. n GALNT3 level of transfected CRC cells was measured. Data are the means ± SD of triplicate determinants (*P < 0.05)