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. 2013 May 22;2(7):675–685. doi: 10.1242/bio.20134507

Fig. 2. hsa-miR29b regulates NSCLC cell proliferation.

Fig. 2.

(A) Real-time PCR analyses of the expression of hsa-miR29b in NSCLC cell lines and non-transformed cell lines. Total RNA was extracted from a non-transformed cell line (Beas2B) or NSCLC cell lines (A549, H157, H661 and H2122) and hsa-miR29b expression was quantified as described in Materials and Methods. RNU6B was used as the internal control for normalization. Data represent mean ± SEM of three separate experiments performed in duplicates. ##P<0.01; versus control (Beas2B). (B) Real-time PCR analyses of the extent of knockdown of hsa-miR29b in Beas2B cells. Beas2B cells were treated with either negative control or synthetic double stranded miR29b precursors. After 48 h, total RNA was extracted and the expression levels of hsa-miR29b were measured as described in Materials and Methods. RNU6B was used as the internal control for normalization. Data represent mean ± SEM of three separate experiments performed in duplicates. ##P<0.01; versus negative control. (C) Clonogenic assays were performed on Beas2B cells treated with miR29b precursors as described in Materials and Methods. Upper panel represents mean ± SEM of the number of colonies counted from two independent and highly reproducible experiments, while representative images were displayed in the lower panel. **P<0.01; versus negative control. NSCLC cells (A549 or H157) were transfected with either empty vector or phsa-miR29b plasmid and cell proliferation rates were determined either by using a clonogenic assay (D) or an MTS assay (E) as described in Materials and Methods. Upper panel represents mean ± SEM from two independent highly reproducible experiments, while representative images were displayed in the lower panel. #P<0.05; ##P<0.01; versus empty vector control. (F) A549 cells were transfected with or without pLNCX-Wnt7a-HA plasmid followed by treatment with miR29b precursors. Cell proliferation rates were later determined using an MTS assay as described in Materials and Methods. Data represent mean ± SEM from three independent highly reproducible experiments. **P<0.01; versus A549+pLNCX-Wnt7a-HA.