Fig. 3

Lamin A/C modulated actin-regulated inflammation. A-B) Representative images and quantitative analysis of F-actin and G-actin levels in RAW264.7 cells cultured on pTi and TNTs before and after 6 h of LPS stimulation. C-D) Subcellular localization of MRTF-A and quantification of the nuclear-to-cytoplasmic ratio in RAW264.7 cells cultured on pTi and TNTs before and after LPS treatment. Lamin B1 was used as an indicator of cytoplasm-nucleus separation. H3 was the loading control for the nuclear fraction, and GAPDH was the loading control for the cytoplasmic fraction. E-F) Representative images and quantitative analysis of F-actin and G-actin levels in lamin A/C-overexpressing macrophages cultured on TNTs and stimulated with LPS. G-H) Subcellular localization of MRTF-A and quantification of the nuclear-to-cytoplasmic ratio in lamin A/C-overexpressing macrophages cultured on TNTs and stimulated with LPS. I) Inflammatory gene expression in lamin A/C-overexpressing and cytochalasin D (Cyto. D) + lamin A/C-overexpressing macrophages after LPS stimulation. Quantification of the F/G-actin ratio was performed on pooled samples from 23–31 single cells in each group from at least 8–12 randomly chosen fields. Western blotting and qRT‒PCR data are presented as the mean ± SD of three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001