Fig. 2: Pharmacological inhibition of TYK2 promotes EMT and invasion in human breast acini and patient-derived TNBC organoids. | Nature Communications

Fig. 2: Pharmacological inhibition of TYK2 promotes EMT and invasion in human breast acini and patient-derived TNBC organoids.

From: Extracellular matrix rigidity controls breast cancer metastasis via TYK2-mediated mechanotransduction

Fig. 2: Pharmacological inhibition of TYK2 promotes EMT and invasion in human breast acini and patient-derived TNBC organoids.The alternative text for this image may have been generated using AI.

a, b Lysates from 3D-PA gels cultured human MCF10A acini overexpressing FLAG-tagged wild-type TYK2 (a) or endogenous TYK2 (b) were subjected to anti-FLAG (a) or anti-TYK2 (b) immunoprecipitation, followed by immunoblot analysis of phosphorylated TYK2 (Tyr1054/1055) and total TYK2 (a) or phospho-tyrosine and TYK2 (b). IgG was used as a negative control, representative of two independent experiments. c, d Human MCF10A acini grown on 3D-PA gels were treated for 5 days with deucravacitinib, AZD1480, or vehicle (DMSO) at the indicated concentrations. Representative bright-field images (c) and quantification of invasive structures (fraction of total) (d) (n = 5 wells per group, three independent experiments). e, f Human MCF10A acini grown on 3D-PA gels were treated for 5 days with zasocitinib or vehicle (DMSO). Representative bright-field images (e) and quantification of invasive structures (f) (n = 5 wells per group, three independent experiments). g, h PDX-derived organoids (PIM025) grown on 3D-PA gels were treated for 5 days with deucravacitinib or vehicle (DMSO). Representative bright-field images (g) and quantification of invasive structures (h) (n = 5 wells per group, three independent experiments). i, j PDX-derived organoids (PIM025) grown on 3D-PA gels were treated for 5 days with zasocitinib or vehicle (DMSO). Representative bright-field images (i) and quantification of invasive structures (j) (n = 5 wells per group, three independent experiments). k Lysates from PDX-derived organoids (PIM025) grown on 3D-PA gels and treated with deucravacitinib (200 nM) or vehicle for 5 days were subjected to anti-TYK2 immunoprecipitation followed by immunoblot analysis of phosphorylated TYK2 (Tyr1054/1055) and total TYK2; E-cadherin, vimentin, and TYK2 were analyzed in input lysates with GAPDH as a loading control, representative of two independent experiments. Data are mean ± SEM, where indicated, quantification is shown for different wells from one representative experiment, and similar results were obtained in independent replicate experiments (see n and number of independent experiments stated for each panel), ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns, not significant. Statistical significance was assessed using one-way ANOVA followed by Dunnett’s multiple-comparisons test. Scale bars: 100 μm (c, e, g, i). Exact P values and source data are provided in the Source Data file.

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