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. 2014 Dec 19;22(8):1275–1286. doi: 10.1038/cdd.2014.209

Figure 2.

Figure 2

ErbB2 and EGFR are necessary for anoikis protection in KPL-4 and SUM149 cells, respectively. (a) KPL-4 cells were transduced with a lentivirus containing either an empty vector (EV) or an ErbB2 shRNA-containing vector (shErbB2), and western blotting analysis confirmed the knockdown. KPL-4 EV and shErbB2 cells were plated on poly-HEMA-coated plates, and caspase activation was quantified at 48 h as previously described. (b) KPL-4 EV and shErbB2 cells were plated at 60 000 cells/well in soft agar as described above. Colonies were stained at 14 days with 0.01% INT-violet and quantified with ImageJ. (c) SUM149 cells were transduced with a lentivirus containing either an empty vector (EV) or an EGFR shRNA-containing vector (shEGFR), and western blotting analysis confirmed the knockdown. SUM149 EV and shEGFR cells were plated on poly-HEMA-coated plates, and caspase activation was quantified at 48 h as previously described. (d) SUM149 EV and shEGFR cells were plated at 100 000 cells/well in soft agar as previously described. Colonies were stained after 30 days using 0.01% INT-violet and quantified with ImageJ. Error bars represent S.E.M.