Journal: Journal of Cellular and Molecular Medicine
Article Title: WTAP ‐Mediated m 6 A Modification of circSMOC1 Accelerates the Tumorigenesis of Non‐Small Cell Lung Cancer by Regulating miR ‐612/ CCL28 Axis
doi: 10.1111/jcmm.70207
Figure Lengend Snippet: MiR‐612 inhibitor abolished circSMOC1 knockdown‐caused CCL28 downregulation in NSCLC. (A) Schematic representation of the binding sites between miR‐612 and WT or Mut CCL28 3′UTR. (B) Analysis of luciferase activities of WT or Mut CCL28 3′UTR after co‐transfection with miR‐612 mimics and WT or Mut CCL28 3′UTR in PC‐9 and 95D cells. (C, D) RT‐qPCR and Western blotting analysis of the expression levels of CCL28 after co‐transfection with sh‐circSMOC1 and miR‐612 inhibitor in PC‐9 and 95D cells. (E, F) Colony formation and transwell analysis of the cell colonies and invasion capabilities after co‐transfection with si‐CCL28 and miR‐612 inhibitor in PC‐9 and 95D cells. Data shown are the mean ± SEM of three experiments. Data shown are the mean ± SEM of three experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.
Article Snippet: The supernatants were resolved in SDS‐PAGE and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore), incubated with anti‐WTAP (10200‐1‐AP, Proteintech), anti‐caspase‐9 (AF6348, Affinity), anti‐cleaved caspase‐9 (AF5240, Affinity), anti‐caspase‐7 (DF6441, Affinity), anti‐cleaved caspase‐7 (AF4023, Affinity), anti‐caspase‐3 (DF6020, Affinity), anti‐cleaved caspase‐3 (AF7022, Affinity), anti‐CCL28 (N3C3, GeneTex) and anti‐GAPDH (AB‐P‐R 001) overnight at 4°C.
Techniques: Knockdown, Binding Assay, Luciferase, Cotransfection, Quantitative RT-PCR, Western Blot, Expressing
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