Journal: Thoracic Cancer

Article Title: MiR ‐429 suppresses proliferation and invasion of breast cancer via inhibiting the Wnt/β ‐catenin signaling pathway

doi: 10.1111/1759-7714.13620

Figure Lengend Snippet: MiR‐429 inhibits the proliferation, migration, and invasion of breast cancer cells. ( a ) RT‐qPCR analysis of miR‐429 in MCF‐10A and human breast cancer cells. ( b ) RT‐qPCR analysis of miR‐429 in transfected cells. ( c ) Colony formation of MDA‐MB‐231 cells after transfection with con and miR‐429. ( d ) EDU assay of MDA‐MB‐231 cells after transfection with con and miR‐429. ( e ) and ( f ) Wound healing assay of MDA‐MB‐231 cells after transfection with con and miR‐429 ( ) MDA‐MB‐231/con, ( ) MDA‐MB‐231/miR‐429. ( g ) Migration and invasion assays of MDA‐MB‐231 cells after transfection with con and miR‐429 ( ) MDA‐MB‐231/con, ( ) MDA‐MB‐231/miR‐429.

Article Snippet: MiR‐429 mimics, miR‐429 negative control, mimic hsa‐miR‐429, inhibitor (inhi)‐miR‐429, inhibitor (inhi)‐FN1, and inhibitor(inhi)LRP6, were purchased from Sigma‐Aldrich Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Migration, Quantitative RT-PCR, Transfection, EdU Assay, Wound Healing Assay

Journal: Thoracic Cancer

Article Title: MiR ‐429 suppresses proliferation and invasion of breast cancer via inhibiting the Wnt/β ‐catenin signaling pathway

doi: 10.1111/1759-7714.13620

Figure Lengend Snippet: MiR‐429 inhibits EMT, cytoskeleton rearrangement and suppresses Wnt/β‐catenin signaling pathway in breast cancer cells. ( a ) Expression of EMT markers tested by western blot, with β‐actin as internal reference. ( b ) Expression of EMT markers evaluated by RT‐qPCR analysis, with β‐actin as internal reference ( ) MDA‐MB‐231/con, ( ) MDA‐MB‐231/miR‐429. ( c ) Expression of EMT markers tested by immunofluorescence. ( d ) Representative images of F‐actin (red) and nucleus (blue) staining in MDA‐MB‐231/con and MDA‐MB‐231/miR‐429 cells; scale bars = 50 μm. ( e ) MiR‐429 suppressed the Wnt signaling TOPflash reporter activity but not the FOPflash reporter. 293T cells were cotransfected with either con plasmid or miR‐429 plasmid, in combination with each pathway luciferase reporter and pRL‐CMV control reporter vectors for 24 hours. A dual‐luciferase assay was performed and results are expressed as fold change ( ) con, ( ) miR‐429. ( f ) The protein levels in the cytoplasm and nucleus of transfected cells. ( g ) The protein levels of Wnt/β‐catenin signaling target genes in MDA‐MB‐231 cells transfected with con or over‐miR‐429.

Article Snippet: MiR‐429 mimics, miR‐429 negative control, mimic hsa‐miR‐429, inhibitor (inhi)‐miR‐429, inhibitor (inhi)‐FN1, and inhibitor(inhi)LRP6, were purchased from Sigma‐Aldrich Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Activity Assay, Plasmid Preparation, Luciferase, Transfection

Journal: Thoracic Cancer

Article Title: MiR ‐429 suppresses proliferation and invasion of breast cancer via inhibiting the Wnt/β ‐catenin signaling pathway

doi: 10.1111/1759-7714.13620

Figure Lengend Snippet: MiR‐429 was able to directly bind to FN1 whose overexpression could restore the effects of miR‐429 on the invasion, migration, EMT and mobility of breast cancer cells in vivo. ( a ) Correlation between miR‐429 and FN1 in breast cancer tissues analyzed by starBase v2.0 ( ) Regression (y = −0.1938x + 8.9021), ( ) r = −0.193, p ‐value = 1.36e −10 . ( b ) Predicted target sites of miR‐429 to 3'‐UTRs of FN1, with the corresponding sequence in the mutated (MU) version. ( c ) The luciferase activity in transfected cells ( ) 293T/con, ( ) 293T/miR‐429. ( d ) RT‐qPCR analysis of FN1 in transfected cells ( ) β‐actin, ( ) FN1. ( e ) Expression of FN1 and EMT markers determined by western blot, with β‐actin as internal reference. ( f and g ) Migration and invasion assays for transfected cells ( ) con + Scr, ( ) miR‐429 + Scr, ( ) con + FN1, ( ) miR‐429 = FN1. ( h ) Immunofluorescence assays for transfected cells. ( i ) The mobility ability of transfected cells ( ) 0H, ( ) 48H.

Article Snippet: MiR‐429 mimics, miR‐429 negative control, mimic hsa‐miR‐429, inhibitor (inhi)‐miR‐429, inhibitor (inhi)‐FN1, and inhibitor(inhi)LRP6, were purchased from Sigma‐Aldrich Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Over Expression, Migration, In Vivo, Sequencing, Luciferase, Activity Assay, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence