Figure 2.

CTR1‐Copper axis displays an important role in AKT activation under pathological conditions. a,b) Ctr1 deletion and counterpart MEFs or CTR1 knockdown and control DLD1 cells were infected with lentivirus encoding PIK3CA‐H1047R, and selected with hygromycin (100 µg mL⁻¹) for 5 days. Resulting cells were subjected for IB analysis. c,d) Cells generated in (b) were subjected to colony formation (c, top panel) and soft agar (d, top panel) assays. The relative colony numbers were normalized and plotted (c, d, bottom panels) (mean ± SD, n = 3; t‐test). e,f) HEK293 cells edited with CRISPR/Cas9 to generate PTEN^−/− and control (sgGFP) cells were infected with tet‐on inducible shRNAs against CTR1, after selected with puromycin (1 µg mL⁻¹) for 72 h, resulting cells were treated with doxycycline (1 µg mL⁻¹) for 48 h and then subjected to IB analysis (e) and colony formation assays (f, top panel). The relative colony numbers were normalized and plotted (f, bottom panel) (mean ± SD, n = 3; t‐test).