Journal: Chinese Medical Journal

Article Title: Effect of down-regulation of let-7c/g on triggering a double-negative feedback loop and promoting restenosis

doi: 10.1097/CM9.0000000000002763

Figure Lengend Snippet: Lin28a interacted with let-7c, let-7g, and miR98, and further regulates the proliferation and migration of VSMCs. (A–E) Let-7c, let-7g, miR98, NC and inhibitor NC oligos were transfected into VSMCs. (A) qRT-PCR validation of successful let-7c, let-7g or miR98 transfections. P <0.05 (let-7c, let-7g, miR98 mimic) vs . NC; P <0.05 (let-7c, let-7g, miR98 inhibitor) vs . inhibitor NC. (B) The EdU assay illustrated the proliferation of VSMCs. VSMCs were labeled with nucleoside analog EdU (red) and nuclei were labeled with Hoechst 33342 dye (blue). The colocalization of EdU (red) and Hoechst 33342 (blue) was shown as pink color, indicative of the proliferative VSMCs. (C) The quantification of the EdU-positive cells. (D) The Transwell assay illustrates the migration of VSMCs. The migrated VSMCs are presented in purple. (E) The quantification of the migration of VSMCs. (F–I) Let-7c, let-7g, miR98, NC and inhibitor NC oligo were transfected into the Lin28a Lenti-virus transfected VSMCs. (F,G) The EdU assay illustrated the proliferation of VSMCs. (H,I) The Transwell assay illustrated the migration of VSMCs. The images of EdU and Transwell assays were taken at magnification ×200 and the scale bar was 75 µm ( n = 5). EdU: 5-ethynyl-2΄-deoxyuridine; miR98: microRNA98; NC: Normal control; qRT-PCR: Quantitative real-time polymerase chain reaction; shRNA: Short-hairpin RNA; VSMCs: Vascular smooth muscle cells.

Article Snippet: DNA was stained with Hoechst 33342 (RiboBio) for 30 min.

Techniques: Migration, Transfection, Quantitative RT-PCR, EdU Assay, Labeling, Transwell Assay, Virus, Real-time Polymerase Chain Reaction, shRNA