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. 2018 Oct 2;20:e00285. doi: 10.1016/j.btre.2018.e00285

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© 2018 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 2 — Stable expression of Gata1 and Fli1 in fibroblasts. MEFs were stably transduced with pMSCV-Hygro-Gata1 or pMSCV-Puro-Fli1 and expression of Gata1 (A) and Fli1 (B) were confirmed by real time RT-PCR, normalised to levels of 18S rRNA and with empty vector (EV) set to 1. Horizontal lines represent the mean of two independent experiments. Nuclear extracts were also prepared from stably transduced cells to assess GATA1 (D) and FLI1 (E) protein levels by Western blot with normalisation to β-actin. Anti-GATA1 N6 (Santa Cruz) and anti-FLI1 C-19 (Santa Cruz) antibodies were used to probe for each protein respectively. Two independent cell lines of EV (lanes 1 and 2) and GATA1/FLI1-transduced MEFs (lanes 3 and 4) were used for Western blots.