Fig. 5

Cell viability test, mRNA transcriptions, and protein changes in response to the mechanical change. A) Cell viability with motionless ZFO, with magnetic field only, and after different action times of ZFO micromotors (30, 60, 90 min) compared to the control group by CCK-8 assay. B) The fluorescent images of Calcein-AM/PI stained A549 cells after different treatments (without ZFO, with ZFO but without rotational magnetic field, and after the action of magnetic ZFO micromotors for 90 min). Scale bar, 100 µm. C) The LDH leakage amount after different treatments (without ZFO, with ZFO but without rotational magnetic field, and after the action of magnetic ZFO micromotors for 90 min). D) The mRNA quantification of the Piezo1 gene after different treatments (without ZFO, with motionless ZFO, with magnetic field only, after the action of ZFO micromotors for 30, 60, and 90 min). E) The mRNA quantification of the ROCK1 gene after different treatments (without ZFO, with ZFO but without rotational magnetic field, after the action of ZFO micromotors for 30, 60, and 90 min). F, G) Protein levels of Piezo1 and ROCK1 upon different treatments (without ZFO, with ZFO but without rotational magnetic field, after action of ZFO micromotors for 30, 60, 90 min) by Western Blot Analysis, with GAPDH (glyceraldehyde-3-phosphate dehydrogenase) as control. H–J) The localization of F-actin in A549 cells. The bright field images (H) and fluorescent images of F-actin in green and Hoechst for nucleus in blue (I) of A549 after different treatments (without ZFO, with ZFO but without rotational magnetic field, and after the action of magnetic ZFO micromotors for 90 min). (J) The enlarged views of F-actin staining. Scale bar: 50 µm (H-I), 6.5 µm (J). K) Statistical changes in cell area of A549. The number of cell statistics for each group is 100. Data are means ± SD of three independent experiments and *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 versus control.