Fig. 4: Suppression of autophagy by mTOR sensitizes to 2DG/DCA. | Nature Communications

Fig. 4: Suppression of autophagy by mTOR sensitizes to 2DG/DCA.

From: mTOR-mediated cancer drug resistance suppresses autophagy and generates a druggable metabolic vulnerability

Fig. 4

a Clonogenic growth assay. H460par cells were pre-treated with 3-methyladenine (3-MA), chloroquine or ULK inhibitor SBI-0206965 for 48 hours before adding 2DG/DCA. H460res cells are shown as control. b, c H460par cells were transfected with a control siRNA (nsi) or siRNAs targeting ATG7. b Western blot. c Clonogenic growth under 2DG/DCA treatment. d–f H460par subclones with indicated CRISPR-engineered ATG7 mutations were analyzed for mTOR signaling, autophagy markers and response to 2DG/DCA treatment. d, Western blot. e, Clonogenic growth. f, Proliferation curves determined by real-time live-cell imaging. Shown is cell culture confluence as mean ± SD, n = 3, FDR q values. g, h H460par clones with CRISPR-induced ATG7 indel mutations were made CDDP-resistant by dose escalation and tested for mTOR-dependent response to 2DG/DCA treatment. g, Flow cytometry analysis for apoptosis (sub-G1). Shown are mean ± SD, n = 3, FDR q values. h Clonogenic growth of CDDP-resistant H460 cells with indicated ATG7 genotype treated with 2DG/DCA ± AZD8055. Shown are representative images.

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