Fig. 3: Hydrogels with high plasticity promote focal adhesion (FA) formation in encapsulated ECs.

a, b Representative immunofluorescent images and quantification of the normalized intensity of integrin β1 (integrin β1 in green, nuclei in blue) (from left to right n = 27, 19, 28 cells from biological triplicate). Scale bar: 50 µm. c, d Protein expression levels of integrin β1 determined by Western blotting of ECs encapsulated in different plastic hydrogels and quantified show increased expression of the integrin of ECs encapsulated in hydrogels with increased network plasticity (n = 4 tests). e, f Representative immunofluorescence images and quantification (n = 30 cells for each group from biological triplicate) of the normalized intensity of p-FAK show increased p-FAK activation of ECs encapsulated in hydrogels with increasing network plasticity (p-FAK in green, nuclei in blue). Scale bar: 50 µm. g Protein expression levels of FAK (h) (n = 6 tests) and p-FAK (i) (n = 3 tests) determined by Western blotting of ECs encapsulated in different plastic hydrogels show increased expression of the integrin of ECs encapsulated in hydrogels with increased network plasticity. j, k Representative immunofluorescent images and quantification of the normalized intensity of p-MLC show enhanced cell contractility of ECs encapsulated in hydrogels with increasing network plasticity (F-actin in red, p-MLC in green, nuclei in blue) (from left to right n = 33, 29, 31 cells for each group from biological triplicate). Scale bar: 25 µm. l Corresponding quantification of the percentage of nuclear localization to overall protein levels (from left to right n = 30, 29, 30 cells for each group from biological triplicate). m, n Protein expression levels of p-MLC determined by Western blotting of ECs encapsulated in different plastic hydrogels (n = 3 tests). All the data are presented as mean values ± SEM. Student’s t-tests are used to assess statistical significance.