Panel A shows fluorescence microscopy images of Acetylated Tubulin and Total Tubulin in cells cultured on Fibronectin for 1 hour, 4 hours, and 24 hours, and on Poly-L-lysine for 1 hour, 4 hours, and 24 hours, with dashed outlines indicating cell boundaries. Panel B shows a superplot of Acetylated/Total Tubulin fluorescence intensity, with the x-axis labeled Fibronectin and Poly-L-lysine at 1 hour, 4 hours, and 24 hours, and the y-axis labeled Acetylated/Total Tubulin fluorescence intensity. Panel C shows fluorescence microscopy images of Acetylated Tubulin and Total Tubulin in Control and 15 percent stretch cells, with dashed outlines indicating cell boundaries. Panel D shows a superplot of Acetylated/Total Tubulin fluorescence intensity, with the x-axis labeled Control and 15 percent stretch, and the y-axis labeled Acetylated/Total Tubulin fluorescence intensity.
Cell spreading induces tubulin acetylation. (A) Representative confocal microscopy images of the acetylated and total α-tubulin fractions of fixed HeLa cells seeded for the indicated times on FN- or PLL-coated glass coverslips. Scale bars, 10 µm. Cell contours are indicated by dashed white curves. (B) SuperPlot showing individual cell measurements (small, light-colored symbols; N∼10 per biological replicate) and the mean value for each independent biological replicate (larger, black-outlined circles; n = 3 replicates). Each color represents a different experimental replicate. A repeated-measures two-way ANOVA test was performed for FN-seeded cells. P values using Fisher’s LSD test are reported in the figure. (C) Representative confocal microscopy images of the acetylated and total α-tubulin fractions of fixed HeLa cells seeded over FN-coated PDMS membranes and subjected to no external forces of 15% mechanical strain. Scale bars, 10 µm. (D) SuperPlot showing individual cell measurements (small, light-colored symbols; N∼10 per biological replicate) and the mean value for each independent biological replicate (larger, black-outlined circles; n = 3). Each color represents a different experimental replicate. A two-sided parametric ratio paired t test was used. The P value is indicated in the plot.