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. 2021 Mar 31;19(3):e3001176. doi: 10.1371/journal.pbio.3001176

Fig 1. Inducible A3A expression reduces the fitness of LUAD cell lines.

Fig 1

(A) A3A mRNA levels in NCI-H358, LXF-289, A549, and A549TP53−/− cell lines transduced with a DOX-inducible A3A cassette at various concentrations of DOX. qRT-PCR and western blot analysis were repeated 2 times. (B) Western blot detection of HA-A3A upon DOX induction. LXF-289, NCI-H358, A549, and A549TP53−/− cells were collected and lysed 72 h posttreatment. Vinculin serves as a loading control, and molecular mass is indicated in kilodaltons. (C) Growth (percentage of growth rate relative to the cells without DOX) for the indicated cell lines after 72 h of DOX treatment measured with AlamarBlue. In red, cells transduced with the inducible A3A cassette, and in black, the parental cell line (no A3A) exposed to the same concentration of DOX. Growth assays were repeated 2 (A549TP53−/−) or 3 times (all other lines). For all graphs, mean and SEM are shown. Uncropped western blots are provided in S1 Raw Images, and numerical data underlying plots are provided in S1 Data. A3A, APOBEC3A; DOX, doxycycline; HA, haemagglutinin; LUAD, lung adenocarcinoma; qRT-PCR, quantitative real-time PCR.