Extended Data 7. Binding of Cu to ULK1 is required for tumorigenesis by oncogenic KRAS^G12D.

a, Mean tumor volume (mm³) ± s.e.m. versus time (days) in mice injected with Ulk1^−/− MEFs stably expressing either HA-ULK1^WT or HA-ULK1^CBM and transformed with KRAS^G12D. n=4 biologically independent animals. Results were compared using a paired, one-tailed Student’s t-test. **, P=0.0095. b, Representative dissected tumors from mice injected with Ulk1^−/− MEFs stably expressing either HA-ULK1^WT (WT) or HA-ULK1^CBM (CBM) and transformed with KRAS^G12D. Scale bar, 100 μm. c, Scatter dot plot with bar at mean tumor weight (g) ± s.e.m. of tumors at endpoint from Ulk1^−/− MEFs stably expressing either WT or CBM and transformed with KRAS^G12D. n=4 biologically independent samples. Results were compared using an unpaired, one-tailed Student’s t-test. *, P=0.0325. n=4. d, Immunoblot detection of T-ULK1, T-ULK2, or β-ACTIN from Kras^G12D/+;Trp53^flox/flox (KP) lung adenocarcinoma cell line #2474 (KP #2474) stably expressing sgRNA against Rosa (−) or Ulk1 and Ulk2 (+). e,f, Scatter dot plot with bar at mean normalized ΔLC3-II/β-ACTIN or ΔP-ATG13/T-ATG13 from Figure 8a,b. n=3 independent experiments. Results were compared using a one-way ANOVA or a two-way ANOVA followed by a Tukey’s multi-comparisons test. e, **, P=0.0013; ***, P=0.0016; ****, P<0.0001; f, *, P=0.0113; **, P=0.0063; ***, P=0.0113. g, Scatter dot plot with bar at mean normalized quantitative PCR (qPCR) expression of Ctr1 mRNA from KP #2474 cells stably expressing sgRNA against Rosa, Ctr1 #2, or Ulk1, Ulk2, and Ctr1 #2. n=1 performed in technical triplicate.