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[Preprint]. 2024 Jun 4:2024.06.04.597454. [Version 1] doi: 10.1101/2024.06.04.597454

Figure 2: Stiffness-dependent vasculogenic mimicry is reduced when pHi is increased on stiff ECM.

Figure 2:

a) Schematic of vasculogenic mimicry (VM) in 2D on stiffening matrix. b) Representative images of H1299 cells stably expressing mCherry-pHluorin pH biosensor plated on soft (0.5% PEGDA) and stiff (4% PEGDA) HA gels and stiff (4% PEGDA) with raised pHi. Images show ratiometric display of pHluorin/mCherry fluorescence ratios. Scale bars: 50 μm. c) Quantification of single-cell pHi data collected as shown in (b) (n=3 biological replicates; n=201 soft, n=237 stiff, n=239 stiff high pHi. Red lines show medians ± IQR). d) Representative images of H1299 cells plated on soft (0.5% PEGDA) and stiff (4% PEGDA) HA gels. Images show differential interference contrast (DIC) and Hoechst stain (DNA, cyan). Scale bars: 100 μm. e) Quantification of cell proliferation across manipulation conditions. (n=3 biological replicates, n=9 per condition. Red lines show means ± SEM).