Fig. 2: Stiffness-dependent vasculogenic mimicry is reduced when pHi is increased on stiff ECM. | Cell Death & Disease

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

From: Intracellular pH dynamics respond to extracellular matrix stiffening and mediate vasculogenic mimicry through β-catenin

Fig. 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 (culture media supplementation with 50 mM sodium bicarbonate). Images show ratiometric display of pHluorin/mCherry fluorescence. 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 and stiff (4% PEGDA) with raised pHi. Images show differential interference contrast (DIC), Hoechst stain (DNA, cyan), and live-cell membrane dye (CellMask, magenta). Scale bars: 100 μm. e Quantification of cell proliferation across manipulation conditions. (N = 3 biological replicates. Red lines show means ± SEM).

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