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. 2013 Aug 15;8(8):e71965. doi: 10.1371/journal.pone.0071965

Figure 6. An SMN-dependent cell proliferation assay in 96-well format.

Figure 6

(A) Automated determination of NIH3T3 cell number in 96-well format. Two-fold serial dilutions of NIH3T3-SMN2low/SMNRNAi cells were plated in eight replicate wells of a 96-well plate. Following fixation and Hoechst staining 4 hours later, cell number was determined by imaging whole wells with an IN Cell Analyzer 2000. (B) Representative IN Cell Analyzer images of NIH3T3-SmnRNAi cells cultured with or without Dox in a 96-well format. (C–D) Analysis of SMN-dependent cell proliferation in NIH3T3-SmnRNAi (C) and NIH3T3-SMN2low/SmnRNAi (D) cells using the 96-well format assay. NIH3T3 cells were cultured for 5 days with or without Dox, and then seeded in six replicate wells of a 96-well plate. Dox-treatment was continued throughout the experiment. Cell number was determined at 4 hours (T0), 4 days and 5 days post-plating. Data are represented as mean and SEM (n = 6; *** = p<0.001; two-way ANOVA).