Fig. 3

Tumor suppressor miR-127-3p is a target of circSDHC in RCC. a. Four miRNAs were predicted as the potential target of circSDHC in CSCD and ENCORI databases. b and c. Relative expression detected by qRT-PCR and gel electrophoresis of circSDHC in 786-O and A498 lysates after RNA pull down with circSDHC specific probe or oligo probe. Expression levels were normalized to oligo probe. GAPDH was used as negative control. d. Relative levels of candidate miRNAs were detected by qRT-PCR after being pull down by circSDHC probe or oligo probe. e. Luciferase reporter assay of 786-O with Luc-vector, Luc-circSDHC or Luc-circSDHC-mutant co-transfected with different candidate miRNAs. f. Relative level of circSDHC was detected by qRT-PCR after being pull down by miR-127-3p probe or oligo probe. g. Cellular localization of miR-127-3p (FAM) and circSDHC (Cy3) detected by FISH. Nuclear was label with DAPI dye. h. circSDHC expression was negatively correlative with miR-127-3p in our own patient cohort (n = 140). i and k. Cell migration and invasion abilities of 786-O and A498 transfected with mimics miR-127-3p or mimics NC. j and l. Cell proliferation ability of 786-O and A498 transfected with mimics miR-127-3p or mimics NC