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. 2020 Aug 7;48(17):9521–9537. doi: 10.1093/nar/gkaa662

Figure 4.

Figure 4.

Ectopic expression of the EIF4A1 HippG370S allele confers Hipp resistance in NIH/3T3 cells. (A) Schematic diagram of MSCV co-expressing His6-eIF4A1 and sheIF4A1 (sh4A1). (B) Western blot analysis of eIF4A1 in the indicated transduced cell lines. Extracts were fractionated on a 4–12% gradient NuPAGE Bis-Tris gel, with electrophoresis performed in MES SDS running buffer as recommended by the manufacturer (Invitrogen). (C) Competition assay of transduced NIH/3T3 cells assessing ability of EIF4A1G370S cDNA to confer Hipp-resistance. Transduced cells (GFP+) were mixed with parental cells (GFP−) and cultured in the presence of 250 nM Hipp for the indicated periods of time. P-values were calculated using the fold changes and a two-way ANOVA followed by Tukey's multiple comparison test. n = 3 ± SD.