Journal: bioRxiv

Article Title: Nascent extracellular matrix converts biomaterial cues into cell fate decisions

doi: 10.64898/2026.01.10.698826

Figure Lengend Snippet: a. Timeline and schematic showing the blocking of cell adhesion to nECM using an integrin β1 (ITGB1) function perturbing antibody for 1 hour pre-embedding and for 7 consecutive days . b. Representative fluorescent images of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (scale bar = 100 μm). c. Quantification of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (low-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 30 ROIs, N = 3; high-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 27 ROIs, N = 3). d. Representative fluorescent images of Sox9 immunofluorescence (dashed line outlines nuclei) of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (scale bar = 10 μm). e. Quantification of Sox9 nucleus-to-cytoplasm (NC) ratio of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (low-Ctrl: n = 138 cells, N = 4; low-ITGB1i: n = 133 cells, N = 4; high-Ctrl: n = 124 cells, N = 4; high-ITGBi: n = 138 cells, N = 4). a-e. N = number of independent experiments ****p < 0.0001, *p < 0.05, error bar = standard deviation, ns: no significant difference by two-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: For perturbation studies, media was supplemented with either an anti-CD44 antibody (DSHB, H4C4, 2.5 μg/mL, day 0-3) or anti-integrin β1 antibody (anti-ITGB1, DSHB, AIIB2, 2.5 μg/mL, day 0-7).

Techniques: Blocking Assay, Cell Culture, Inhibition, Modification, Immunofluorescence, Standard Deviation

Journal: bioRxiv

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

doi: 10.1101/2025.10.30.681251

Figure Lengend Snippet: a) Bright field images of the human MCF10A acini that were grown on 3D-PA gels and treated with β1 integrin AIIB2 blocking antibody or vehicle control at indicated concentrations for 5 days. Scale bar represents 100μm. b) Invasive structures of the β1-integrin blocking antibody AIIB2 or vehicle control treated human MCF10A acini grown on 3D-PA gels for 5 days were quantified and represented as fraction of total structures. Dots represent individual fields. ****p < 0.0001; ns, not significant. c) Bright field images of the TYK2 overexpressed human MCF10A acini that were grown on 3D-PA gels and treated with the β1 integrin blocking antibody AIIB2, FAK inhibitor VS-4718 (0.2μM) or vehicle control (DMSO) at indicated concentrations for 5 days. Scale bar represents 100μm. d) Invasive structures of the β1-integrin blocking antibody AIIB2, FAK inhibitor VS-4718 (0.2μM) or vehicle control (DMSO) the TYK2 overexpressed human MCF10A acini grown on 3D-PA gels for 5 days were quantified and represented as fraction of total structures. Dots represent individual fields. *p < 0.05; * * p < 0.01; ****p < 0.0001; ns, not significant. e) Immunoblot analysis of pFAK, total FAK and TYK2 expression in lysates from the TYK2 overexpressed human MCF10A treated with the β1-integrin blocking antibody AIIB2, FAK inhibitor VS-4718 (0.2μM) or vehicle control (DMSO) that were grown on 3D-PA gels. GAPDH is used as a loading control. f) MCF10A acini overexpressing FLAG-TYK2 WT and treated with the β1-integrin blocking antibody AIIB2, FAK inhibitor VS-4718 (0.2μM) or vehicle control (DMSO) that were grown on 3D-PA gels. Lysates were subjected to anti-FLAG immunoprecipitation were analysed with immunoblotting. g) MCF10A acini overexpressing FLAG-TYK2 WT and treated with the β1-integrin blocking antibody AIIB2 or vehicle control that were grown on 3D-PA gels. and then immunostained for TYK2 (red) and DAPI (blue). Scale bar represents 25μm. h) Plots of DAPI and TYK2 intensity profiles plotted as a function of distance across tissue sections versus their pixel intensities in human MCF10A acini overexpressing FLAG-tagged TYK2 and treated with the β1-integrin blocking antibody AIIB2 or vehicle control that were grown on 3D-PA gels.

Article Snippet: Antibodies used in this study included TYK2 specific for human, Sigma-Aldrich, HPA005157; TYK2, GeneTex, GTX61449; pTyr1054/1055 TYK2, Cell Signaling Technology, #68790; Phospho-Tyrosine, Cell Signaling Technology, # 9411; TWIST, Santa Cruz Biotechnology, sc-81417; G3BP2, Sigma-Aldrich, HPA018425; E-cadherin, BD Biosciences,610181;Vimentin, Cell Signaling Technology, #5741; Fibronectin, Sigma-Aldrich, F3548; STAT3, Cell Signaling Technology, #9139; pTyr705 STAT3, Cell Signaling Technology, #9145; STAT1, Cell Signaling Technology, #9172; pTyr701 STAT1, Cell Signaling Technology, #9167; STAT5, Cell Signaling Technology, #57580; pTyr694 STAT5, Cell Signaling Technology, #4322; EPHA2, Cell Signaling Technology, #6997; pS897 EPHA2, Cell Signaling Technology, #6347; LYN, Cell Signaling Technology, #2796; pY416 SFK, Cell Signaling Technology, #2101; IFNAR1, Abcam, ab45172; FLAG, Cell Signaling Technology, #14793; Rabbit IgG, Cell Signaling Technology, #2729; FAK, Cell Signaling Technology, #13009; pTyr397 FAK, Invitrogen, # 44-625G; AIIB2 β1-integrin-blocking antibody, Developmental Studies Hybridoma Bank, AB_528306; Laminin V, kind gift from M. Aumailley, University of Cologne, Germany; KRT5, Invitrogen, # PA5-32465; Ki-67, Invitrogen, # MA5-14520; GAPDH, GeneTex, GTX100118.

Techniques: Blocking Assay, Control, Western Blot, Expressing, Immunoprecipitation