Figure 4
From: Glucocorticoid therapy regulates podocyte motility by inhibition of Rac1
Effects of Gc and puromycin aminonucleoside (PAN) exposure on podocyte motility. Microarray analysis of Gc-regulated genes from wild-type podocytes suggested Gc exposure affected podocyte motility. In order to test this, live cell imaging of wild type podocytes was performed for a 24 hour period, beginning 24 hours following treatment. Subsequently, manual cell tracking was performed using the MTrackJ plugin on ImageJ software to allow analysis of podocyte speed and cell directional persistence. Each experiment consisted of tracking 120 cells per condition. The experiment was performed 3 times. (a) Quantifies mean podocyte speed over the 24 hour period. (b) Quantifies podocyte directional persistence. Rose plots were created for each condition to visualize the path travelled for 20 cells (the path of each cell is marked with a different colour). The x- and y-axis refer to x,y coordinates of each cell over time generated by the cell tracking software: (c) vehicle-treated; (d) Gc-treated; (e) PAN-treated; (f) PAN + Gc-treated. (g) is a rose plot comparing the movement of 10 PAN-treated podocytes (red) compared with 10 podocytes treated with PAN + Gc (black) over the 24 hour period. Results were compared to vehicle-treated and analysed by one-way ANOVA followed by Dunnett’s multiple comparisons test. ** = adjusted p value 0.091; *** = adjusted p value 0.0003; **** = adjusted p value < 0.0001. N.S. = not significant. Error bars represent standard error of the mean.
