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. Author manuscript; available in PMC: 2022 May 12.
Published in final edited form as: Science. 2021 Nov 11;374(6569):848–856. doi: 10.1126/science.abh3602

Fig. 2. Effect of supramolecular motion on hNPCs signaling in vitro.

Fig. 2.

(A) Molecular graphics representation of an IKVAV PA nanofiber indicating the chemical structure and location of DPH used as a probe in fluorescence depolarization measurements (top); bar graph of fluorescence anisotropy of IKVAV PA solutions (error bars correspond to 3 independent experiments; n.s. no significant, ***P<0.0001, one-way ANOVA with Bonferroni). (B) Chemical structure of the IKVAV peptide sequence highlighting the K residue probed by NMR (top); bar graphs of the K relaxation time for the different IKVAV PAs investigated (error bars correspond to 3 runs per condition; ***P<0.0001 vs IKVAV PA1, #P<0.05, ###P<0.0001 vs IKVAV PA2 and +P<0.05, +++P<0.0001 vs IKVAV PA5, one-way ANOVA with Bonferroni). (C) Differentiation conditions used for hNPCs. (D) Representative micrographs of hNPCs treated with IKVAV PA1, PA2, PA4 and PA5; NESTIN-stem cells (red), ITGB1-receptor (green), and DAPI-nuclei (blue). (E) WB results of ITGB1, p-FAK, FAK, ILK, and TUJ-1 in hNPCs treated with laminin and the various IKVAV PAs. (F) Representative confocal micrographs of hNPCs treated with IKVAV PA1, PA2, PA4 and PA5; NESTIN-stem cells (red), SOX-2-stem cells (green), TUJ-1-neurons (white), and DAPI-nuclei (blue). (G, H) Bar graphs of the percentage of SOX-2+ and NESTIN+-stem cells (G) and TUJ-1+ cells (H) treated with the various IKVAV PAs (error bars correspond to 3 independent differentiations; **P<0.01, ***P<0.001 vs IKVAV PA2 and ##P<0.01, ###P<0.001 vs IKVAV PA5, one-way ANOVA with Bonferroni). (I) Fluorescence anisotropy (left) and K residue relaxation times (right) obtained for IKVAV PA2 nanofibers in the absence (No Ca2+) or presence (Ca2+) of calcium ions (***P<0.001, student’s t-test). (J) WB results of ITGB1, p-FAK, FAK, ILK, TUJ-1 in hNPCs treated with IKVAV PA2 in the absence (−) or presence (+) of Ca2+. Scale bars: (D) 10 μm, (F) 100 μm.