Figure 2. An in vivo shRNA screen for modulators of doxorubicin response.

(A) A diagram depicting the in vivo screening strategy. Pooled shRNAs targeting Bcl-2 family members were retrovirally transduced into Eμ-MYC p19^Arf−/− lymphomas. shRNA composition was measured either following the presentation of palpable tumor burden or following tumor relapse after treatment with 8mg/kg doxorubicin. (B) A comparison of the coefficient of variation (CV) for hairpins targeting the Bcl-2 family in cell culture versus in vivo screens using a Luminex measurement methodology. (C) An analysis of the distribution of shRNA CV values for a large in vitro versus in vivo screening data set generated by high-throughput sequencing. The vertical line represents the CV threshold used to filter in vivo data. The arrows denote averages for validated hairpins relative to all hairpins (D) Bcl-2 family “hits” following filtering for CV and enrichment criteria and separated by anatomical niche. The asterisk denotes a Bcl-w shRNA that scored as enriched in the thymus, but was excluded due to the lack of Bcl-w expression in this context.