Fig. 4: SOX2-ZATAC#20 efficiently degrades SOX2 and inhibits cancer cell proliferation. | Nature Communications

Fig. 4: SOX2-ZATAC#20 efficiently degrades SOX2 and inhibits cancer cell proliferation.

From: Identification of a non-inhibitory aptameric ligand to CRL2ZYG11B E3 ligase for targeted protein degradation

Fig. 4

a Schematic of SOX2 degradation mediated by SOX2-ZATACs. Created with BioRender.com. b Immunoblotting analysis of A549 cells treated with SOX2-ZATACs with different lengths of double-stranded flexible linkers for 24 h. c Immunoblotting analysis of A549 cells treated with scramble control (Scr-ZATAC-2#20) or SOX2-ZATAC#20 for 24 h. d Immunoblotting analysis of A549 cells treated with SOX2-ZATAC#20 (500 nM) for the specified time points. e Co-immunoprecipitation assays to evaluate the interaction between endogenous SOX2 and ZYG11B in the presence or absence of SOX2-ZATAC#20 (500 nM) in MG132-treated (20 µM) A549 cells. f, g Immunoblotting analysis of A549 cells treated with SOX2-ZATAC#20 (500 nM), together with or without CQ (10 µM) or MG132 (20 µM) for 24 h. h Immunoblotting analysis of A549 cells treated with SOX2-ZATAC#20 (500 nM) and varying concentrations of Apt#Z6. i Immunoblotting analysis of A549 cells transfected with ZYG11B-targeted siRNAs, followed by treating with SOX2-ZATAC#20 (500 nM) for 24 h. j, k A549 cells were treated with PBS, Apt#SOX2 (500 nM), or SOX2-ZATAC#20 (500 nM) for 24 h, and were subjected to EdU incorporation assays (j). The percentage of EdU-positive cells was quantified (k). Scale bar: 100 µm. l A549 cells were treated with PBS, Apt#SOX2 (500 nM), or SOX2-ZATAC#20 (500 nM) for specified time points, and viable cells were evaluated using CCK-8 assays. m, n A549 cells were treated with PBS, Apt#SOX2 (500 nM), or SOX2-ZATAC#20 (500 nM) for two weeks and then subjected to crystal violet staining. The colonies were imaged (m) and quantified (n). o, p Representative 3D tumor spheroidic image (o) and quantification (p) of A549 cells treated with PBS, Apt#SOX2 (500 nM), or SOX2-ZATAC#20 (500 nM). Scale bar, 65 µm. The averages of n = 5 (k), n = 4 (l), n =  3 (n, p) biological replicates are shown. Data are shown as the mean ± SD. Statistical significance in (k, l, n, p) was assessed using one-way ANOVA with multiple comparisons. The data shown in (b–j, m, o) are representative of three independent experiments.

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