Fig. 5: CTSD deficiency induces autophagy in short-term starved tumor cells. | Nature Communications

Fig. 5: CTSD deficiency induces autophagy in short-term starved tumor cells.

From: Cathepsin D deficiency in mammary epithelium transiently stalls breast cancer by interference with mTORC1 signaling

Fig. 5

a Analysis of LC3-I, LC3-II, and p62 protein levels by Western blot (with TUBA as loading control) in Ctsd+/+ and Ctsd−/− PyMT cells cultured for 7–10 days in 10% FCS or 1% FCS medium. Representative of 4 independent experiments. b Quantification of LC3 Western blots as in a, plotted as LC3-II/LC3-I ratio relative to 10% FCS Ctsd+/+ (n = 4 independent experiments; two-sided one-sample and two-sample t-test). Relative Lc3a (c) and p62 (d) expression determined by RT-PCR in 10% FCS and 1% FCS Ctsd+/+ and Ctsd−/− PyMT cells (n = 3 independent experiments; two-sided two-sample t-test). e Pictures of 10% FCS and 1% FCS Ctsd+/+ and Ctsd−/− PyMT cells stained for LC3 (green), LAMP1 (red), and DNA (blue). Zoom shows enlargement of indicated image area in merge+DNA. Bars, 10 μm. Representative of 3 independent experiments. f Principle component analysis (PCA) of targeted metabolomics data from 1% FCS Ctsd+/+ and Ctsd−/− PyMT cells (n = 3 independent experiments). g Clustering analysis of the relative abundance of all 20 canonical amino acids and cystine in 1% FCS Ctsd+/+ and Ctsd−/− PyMT cells shown as heatmap (n = 3 independent experiments). Bar charts show all data points with mean + SD and p-value. Source data are provided as a Source Data file.

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