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. 2014 Sep 22;9(2):355–364. doi: 10.1016/j.molonc.2014.09.005

Figure 1.

Figure 1

MCF10A 3D culture‐based screening for genes coamplified with ERBB2 and identification of RARA as a gene inducing filled‐lumen structures. (A) Overview of the screening system. Fifty‐two genes on the 17q12–21 amplicon including wild type ERBB2 were selected as screening targets. We divided full‐length cDNAs corresponding to those genes into ten subgroups and introduced them into MCF10A/Tet‐on/TRE‐ERBB2VE/Eco cells using retroviruses. The cells were embedded in Matrigel and allowed to proliferate. (B) Selection of transformed gene subgroups using a 3D culture system. Cells were infected with retrovirus mixture for each subgroup, embedded in Matrigel and cultured for 15 days. In the bottom panels, ERBB2VE was induced with Dox at day 4. Scale bars represent 200 μm. Magnified pictures are provided in Figure S2. (C) Activity of RARA in both ERBB2VE (−) and (+) backgrounds. MCF10A/Tet‐on/TRE‐ERBB2VE/Eco cells were transfected with mock (left panels) or RARA (right panels) retroviruses and were cultured on Matrigel for 14 days. In the right panels, ERBB2VE was induced by the addition of Dox at day 4. Arrows indicate raptured structures. Scale bars represent 200 μm.