Fig. 3: DSS1 enhances ccRCC cell migration and invasion via autophagy inhibition. | Nature Communications

Fig. 3: DSS1 enhances ccRCC cell migration and invasion via autophagy inhibition.

From: DSS1 inhibits autophagy to activate epithelial-mesenchymal transition in a pro-metastatic niche of renal cell carcinoma

Fig. 3

a, b Transwell migration and Matrigel invasion assays in ACHN and Caki-1 cells treated with shNC, shDSS1, pcDNA3.1, DSS1 plasmids, with or without CQ (Chloroquine, representative images; scale bar: 100 μm). Quantification of migrated/invaded cells (error bar: mean ± SD; two-tailed Welch’s t-test; n = 3 independent experiments). c Morphology images of ccRCC cells between DSS1 knockdown (pebble-shaped) and negative control (spindle-shaped; scale bar: 100 μm; error bar: mean ± SD; two-tailed Welch’s t-test). The average cell aspect ratio was determined from 100 randomly selected cells per group per experiment (n = 3 independent experiments). d, e Immunoblotting (IB) of autophagy markers (LC3-II, p62) and EMT proteins (E-cadherin, N-cadherin, Vimentin) in siNC- and siDSS1-treated (with or without CQ, 0.02% DMSO as control) cells. Endogenous control: β-actin. Densitometry quantification (e, n = 3 independent experiments, error bar: mean ± SD, two-tailed Welch’s t-test). The samples derived from the same experiment were run on parallel gels, with each gel probed for a different antibody. f RT-qPCR analysis of MAP1LC3B and SQSTM1 mRNA levels in siDSS1 vs. siNC cells (normalized to GAPDH; n = 3 independent experiments; error bar: mean ± SD; two-tailed Welch’s t-test). Statistics are provided in the source data. Source data are provided as a Source Data file.

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