Increasing mGluR1 activity has no effect on measures of oncogenesis in MCF10A cells.

<p><b>A. <i>GRM1</i> expression.</b> RNA lysates from stable transfectants of <i>GRM1</i>-MCF10AT1 or control <i>LACZ</i> cells were isolated and subjected to qPCR as described in Materials/Subjects and Methods. <i>GRM1</i> is expressed at a level approximately 1.8×10⁴ times higher in GRM1-transduced MCF10A cells than in <i>LacZ</i> control infected cells. <b>B. mGluR1 expression.</b> Protein lysate (30 µg) was loaded in each lane. The membrane was stripped and reprobed for tubulin antibody to show equal loadings. (<b><i>LacZ</i></b> = pLenti-LacZ; <b><i>GRM1</i></b> = pLenti-<i>GRM1</i>). <b>C. Effect of mGluR1 overexpression on proliferation.</b> Lentivirus-driven mGluR1 expression had no effect on the proliferation of MCF10AT1 cells compared to the control as measured by MTT assay. For MTT assay, 1×10⁴ cells were plated and cell proliferation was analyzed on day 3. <b>D. Effect of mGluR1 expression on MCF10A migration.</b> <i>GRM1</i> overexpression in MCF10AT1 cells had no effect on MCF10A migration compared to the control <i>LacZ-</i>transduced cells. <b>E. Effect of mGluR1 expression on anchorage-independent colony formation.</b> MCF10A cells were transduced with either with pLenti-<i>GRM1</i> or pLenti-<i>LacZ</i> and plated in soft agar as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0081126#s2" target="_blank">Materials and Methods</a>. <i>GRM1</i> overexpression had no effect on the number of colonies on soft agar compared to the control <i>LacZ</i> cells. All experimenbts were performed in triplicate and results are recorded as the mean ± SEM of three experiments, where *p<0.05 compared to control.