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Figure 4. Characterization of 231_sEVs released from cells grown on different substrates. Representative AFM images (A), scatterplots and boxplots obtained from the AFM image analysis (B-C), and NTA graphs of fresh 231_sEVs derived from cells plated on different stiffness substrates. The lower and upper boundaries of the box represent Q1 (25 percentile) and Q3 (75 percentile) of the data, respectively; the horizontal bar inside the box represents the median of the data; NTA results confirmed that 231_sEVs released from cells grown on soft PDMS are more abundant and bigger in size compared to the other conditions. Significance of data differences was established via the Anova Kruskal-Wallis test (^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001, respectively). Representative experiments of three independent data sets. AFM: Atomic force microscopy; NTA: nanoparticle tracking analysis; PDMS: polydimethylsiloxane; sEVs: small extracellular vesicles.