Figure 7. Knockdown of ZFX targets DIS3L and UBE2J1 impairs growth of human DAOY medulloblastoma cells in vitro.

(A) Expression of DIS3L and UBE2J1 in DAOY cells after ZFX knockdown. Shown are DIS3L and UBE2J1 expression levels by qPCR 4 days after transduction with lentivirus coding for ZFX-targeting (H2, H3, H4) or scrambled control (SCR) shRNA (mean ± SD of triplicate parallel cultures; representative of three independent experiments).

(B) Expression levels of Dis3L and Ube2j1 in bulk epidermis and hair follicles from shaved lower dorsal skin 3 days after topical Tmx treatment in Ptch1, Ptch1-Zfx, and Cre⁻ (Ctrl) mice (as described in Figure 1). Shown are normalized expression levels relative to Ctrl skin as determined by qPCR (mean ± SD of 2–4 mice; representative of four independent experiments).

(C) DAOY human MB cells were transduced in vitro with Sigma MISSION lentiviruses encoding shRNAs targeting DIS3L, UBE2J1, or a non-mammalian control gene (NonM). Shown are projected cell numbers for untreated DAOY cells versus cells transduced with NonM control virus or 3–4 shRNA lentiviruses targeting DIS3L (SH1–SH4) or UBE2J1 (SH3–SH5) (mean + SD of triplicate parallel cultures; beginning 2 days post- puromycin selection and 4 days post-transduction).

(D) Expression of DIS3L and UBE2J1 in DAOY cells after shRNA knockdown. Shown are DIS3L and UBE2J1 expression levels by qPCR in DAOY cells 5 days after transduction with Sigma MISSION lentiviruses coding for DIS3L- or UBE2J1-targeting shRNA, or for NonM control shRNA (mean ± SD of triplicate reactions; normalized to expression in untreated DAOY cells).