Fig 1: miR-138-5p and TMEM40 expression levels are negatively correlated in ccRCC patient tissues samples. Quantitative real-time PCR analysis for (A) miR-138-5p and (B) TMEM40 in 36 paired ccRCC and adjacent normal tissues. (C) The Pearson’s correlation analysis for the relationship between miR-138-5p levels and TMEM40 mRNA levels in ccRCC tissues.(D and E) Western blot analysis for TMEM40 expression in 8 paired ccRCC and adjacent normal tissues. T1-8 presents tumor tissues and N1-8 presents paired normal tissues.
Fig 2: TMEM40 was the downstream target of miR-138-5p in ccRCC. (A) miR-138-5p and its putative binding sequence in the 3’-UTR of TMEM40. Mutated TMEM40 3’-UTR was generated in the complementary site for the seed region of miR-138-5p. (B) miR-138-5p overexpression significantly suppressed the luciferase activity of wild type TMEM40 3’-UTR but not mutated 3’-UTR of TMEM40. (C) The mRNA level of TMEM40 and (D) the protein level of TMEM40 were significantly inhibited in 786-O cells after miR-138-5p overexpression. (E) The mRNA level of TMEM40 and (F) the protein level of TMEM40 were significantly suppressed in ACHN cells after miR-138-5p overexpression. **p< 0.01, ***p< 0.001, compared with miR-NC; NC, negative control.
Fig 3: The ability of miR-138-5p and TMEM40 in cell migration was assessed by wound healing assay. (A) Representative cell migratory distances images in 786-O and ACHN cell lines. Statistical analysis of wound healing rate in (B) 786-O and (C) ACHN cells after transfection with miR-138-5p mimic or miR-NC and siTMEM40 or siNC; **p< 0.01, compared with miR-NC; ##p< 0.01, ###p< 0.001, compared with siNC; NC, negative control.
Fig 4: The ability of miR-138-5p and TMEM40 in cell invasion was assessed by transwell assay. (A) Representative cell invasive images in 786-O and ACHN cell lines. Statistical analysis of invasive cell number in (B) 786-O and (C) ACHN cells after transfection with miR-138-5p mimic or miR-NC and siTMEM40 or siNC; **p< 0.01, ***p< 0.001, compared with miR-NC; ##p< 0.01, compared with siNC; NC, negative control.
Fig 5: The ability of miR-138-5p and TMEM40 in cell proliferation was assessed by CCK-8 assay. (A) 786-O cells and (B) ACHN cells were transfected with miR-138-5p mimic or miR-NC and underwent CCK-8 assay. (C) 786-O cells and (D) ACHN cells were transfected with siTMEM40 or siNCand underwent CCK-8 assay. Colony formation was measured in ACHN cells transfected with (E) miR-138-5p mimic or transfected with (F) siTMEM40. ***p< 0.001, compared with miR-NC; ##p< 0.01, ###p< 0.001, compared with siNC; NC, negative control; OD, optical density; CCK8, Cell Counting Kit 8.
Supplier Page from Abcam for Anti-TMEM40 antibody