Panel A shows fluorescence microscopy images of GST-C1a-PKD, TGN46, and merged images 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. Panel B shows a superplot of Golgi Diacylglycerol sensor levels (arbitrary units), with the x-axis labeled Fibronectin and Poly-L-lysine at 1 hour, 4 hours, and 24 hours, and the y-axis labeled Golgi Diacylglycerol sensor levels (arbitrary units). Panel C shows fluorescence microscopy images of GST-C1a-PKD, GM130, and merged images in cells cultured on 2 kilopascals, 30 kilopascals, and Glass. Panel D shows a superplot of Golgi Diacylglycerol sensor levels (arbitrary units), with the x-axis labeled 2 kilopascals, 30 kilopascals, and Glass, and the y-axis labeled Golgi Diacylglycerol sensor levels (arbitrary units). Panel E shows fluorescence microscopy images of GST-C1a-PKD, TGN46, and merged images in Dimethyl sulfoxide and Tubacin treated cells. Panel F shows a superplot of Golgi Diacylglycerol sensor levels (arbitrary units), with the x-axis labeled Dimethyl sulfoxide and Tubacin, and the y-axis labeled Golgi Diacylglycerol sensor levels (arbitrary units).
Mechanical cues alter Golgi DAG content. (A) Representative confocal microscopy images of HeLa cells transfected with the GST-C1a-PKD DAG sensor, seeded over FN or PLL for the indicated times, fixed, and immunostained for GST and TGN46. Scale bars, 10 µm. (B) SuperPlot (N∼10 cells per biological replicate; n = 3 biological replicates) of the quantification of A. A repeated-measures two-way ANOVA test was performed. P values using Tukey’s post hoc multiple comparison test are reported. (C) Representative images of HeLa cells transfected with the GST-C1a-PKD DAG sensor, seeded on FN-coated glass coverslips, stiff (30 kPa) or soft (2 kPa) PAA gels, fixed, and immunostained for GST and GM130. (D) SuperPlot showing quantification of C. A repeated-measures one-way ANOVA test was performed using Tukey’s post hoc multiple comparison test (n = 3 replicates; N ≥ 5 cells per replicate). (E) Representative confocal microscopy images of HeLa cells transfected with the GST-C1a-PKD plasmid, subjected to a treatment with 10 µM Tubacin (or DMSO), fixed, and immunostained for GST and TGN46. Scale bars, 10 µm. (F) SuperPlot (N∼10 per biological replicate; n = 3 biological replicates) showing quantification of E. A two-sided parametric ratio paired t test was used. The P value is indicated in the plot.