(A) Schema describing DOX treatment of cells (described in Fig. 2, C to E) and transduction of these cells to either express sgRNA targeting ARNT1 (sgARNT) or non-targeting control (sgCon).
(B–D) Viable cell counts (B), immunoblot analysis (C), and representative photomicrographs (D) of cells (described in Fig. 6A) expressing the indicated sgRNA. In (B), “Days” indicates time after withdrawal of DOX. Images in (D) were recorded 15 days after DOX withdrawal. In (B) p-values were calculated using Students t-test and p<0.05 was considered significant [*].
(E and F) Crystal violet staining (E) and immunoblots (F) of 786-O cells infected to exogenously produce the indicated ARNT1 shRNAs or a control shRNA (Con). In (E), cells were treated with the indicated amounts of JQ-EZ-05 for 10 days. In (F), SE = short exposure and LE = long exposure.
(G and H) Crystal violet staining (G) and immunoblots (H) of 786-O cells infected with lentiviruses encoding either ARNT1 sgRNA or a control sgRNA. In (G), cells were treated with the indicated amounts of JQ-EZ-05 for 10 days. NDRG1 was included in (F) and (H) as a HIF2α-responsive gene product.