Fig. 5: SMYD5 deletion sensitizes cancer cells to mTOR blockade.

a Schematic of drug screen in the control and SMYD5 KO1 Huh7 cells. b Scatter plot of the fitness of SMYD5 KO1 Huh7 cells compared to the NC Huh7 cells under the treatment of individual drugs for 4 days. Data presented as the relative growth rate of SMYD5 KO1/NC cells. Four hits caused more than 25% growth retardation of SMYD5 KO cells compared to the control cells were in red. Data were presented as the mean from duplicated experiments. c Proliferation analyses of the control and SMYD5-depleted (KO1) Huh7, HeLa, SNU449 and HepG2 cells under the treatment of Torin1 at the indicated concentrations. Experiments were performed three times. d WB analyses of newly synthesized proteins in the NC and SMYD5 KO1 Huh7 cell lines with or without Torin1 treatment by puromycin labeling approach. e WB analyses of newly synthesized proteins in the WT, RPL40 K22R KI, and RPL40 K22R KI with further knockdown of SMYD5 HEK293T cell lines with or without Torin1 treatment by puromycin labeling approach. f Scatterplots of ribo-seq depicting the changes between control and KO cells in transcription level (x-axis) and translation level (y-axis) with 50 nM Torin1 treatment for 12 hr. The red and blue dots represent genes with stable mRNA levels but increase or decrease in RPF levels. FC cutoff is 1.5. g Volcanoplots of ribo-seq representing the TE changes. Genes with FC > 1.5 or < 1/1.5 P adj < 0.05 were colored. Average TE is between NC and SMYD5 KO1 cells, allowing for an evaluation of whether genes are highly/lowly translated. h Footprint of ribo-seq across normalized CDS regions under Torin1 treatment, normalized by mitochondrial RPF.