Fig. 3: Inhibition of Dnmt1 eliminates the LPS-stimulated cellular stiffening.

A A schematic diagram of the nanoindentation test to measure the cell stiffness. B, C The cellular stiffness measured by nanoindentation. In (B), RAW 264.7 cells were sequentially treated with 5-Aza/DMSO and LPS/PBS. Each dot means a representative cell from five independent experiments. In (C), peritoneal macrophages from Dnmt1^KO and Dnmt1^fl/fl mice were treated with LPS/PBS. Each dot means a representative cell from 3 to 5 mice. D A schematic diagram of the micropipette aspiration test to measure the cell viscoelasticity. E Representative brightfield images of RAW 264.7 cells immediately before (0 s) and after (1 s) partial aspiration into a micropipette. The cells were treated as in (B). F The viscoelastic parameters (k₁, k₂ and μ) of RAW 264.7 cells from (E). Each dot represents a single cell. Data were obtained from 3–5 independent experiments. G Immunofluorescence staining of cytoskeletons in RAW264.7 cells treated with cytoskeleton depolymerization compounds (acrylamide, cytochalasin, or nocodazole), respectively. Scale bar, 20 μm. H Representative brightfield images of RAW 264.7 cells immediately before (0 s) and after (1 s) partial aspiration into a micropipette. The cells were sequentially treated with cytoskeleton depolymerization compounds, 5-Aza/DMSO, and LPS/PBS. I The viscoelastic parameters (k₁, k₂ and μ) of RAW 264.7 cells from (H). Each dot represents a single cell. Data were obtained from 3-5 independent experiments and are presented as mean ± SEM. *P < 0.05 by one-way ANOVA followed by Turkey’s post hoc test.