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. 2024 May 17;15:3812. doi: 10.1038/s41467-024-48088-5

Fig. 3. ELASTomics analysis of cellular senescence.

Fig. 3

a Fluorescence images of nanopore-electroporated TIG-1 cells at different population doubling levels (PDL) = 42 and 59. FITC-BSA (green) was imported into cells by nanopore-electroporation. Nuclei were stained by Hoechst 33342 (blue). Similar results were observed in independent duplicate experiments. Scale bar, 50 μm. b Centered log ratio of 4 kDa DTD in TIG-1 cells at PDL = 42 and 59. Dots indicate individual cells. c-e UMAP of relatively young (PDL = 42) and senescent (PDL = 59) TIG-1 cells. The colors respectively indicate the applied voltages (c), PDLs (d), and centered log ratio of 4 kDa DTD (e). f The coefficients of correlation between the centered log ratio of 4 kDa DTD and expression of individual genes. Red points are the genes with Pearson’s |r | > 0.15. g The coefficients of variables (s) and the coefficients of correlation between DTD counts and gene expression. Red points are genes with Pearson’s r > 0.3 or < −0.2 (P < 0.001) and |s | > 0.1. h, i Comparison of bulk TIG-1 cells at different PDLs (PDL = 33, 44, 52, and 65) with the expression levels of KLF2 (h) and RRAD (i) (n = 3 biologically independent samples). J-l Fluorescence intensity of imported FITC-BSA by nanopore-electroporation to TIG-1 cells with different PDLs (PDL = 33 n = 11,008; PDL = 44 n = 11,050; PDL = 52 n = 11,171; PDL = 65 n = 9365 independent cells) (j), with or without the suppression of RRAD by siRNA (PDL = 40 with siNegative n = 9711; PDL = 40 with siRRAD n = 11,012; PDL = 56 with siNegative n = 10,636; PDL = 56 with siRRAD n = 4894 independent cells) (k), and treated with vehicle or 2-deoxy-D-glucose (2-DG) (PDL = 40 with 0 mM 2-DG n = 10,931; PDL = 40 with 1 mM 2-DG n = 11,076; PDL = 60 with 0 mM 2-DG n = 2861; PDL = 60 with 1 mM 2-DG n = 6083 independent cells) (l). In the boxplots (j–l), the center lines are the median, the box indicates the first and third quartiles, whiskers are the minimum and the maximum excluding outliers, and dots are outliers. Similar results were observed in independent duplicate experiments. The P values (p) are indicated in the graph (h-l: Tukey’s t-test). Source data are provided as a Source Data file.