Pitstop 2 Is a Potent Inhibitor of Clathrin-Independent Endocytosis (original) (raw)
Figure 2
Pitstop 2 inhibits clathrin-independent endocytosis in a dose-dependent manner.
(A) Hela cells were preincubated with DMSO (0.1%) or different doses of pitstop 2 ranging from 5 µM to 30 µM. Cells were then allowed to internalize Alexa594-Transferrin and antibodies to MHCI in the presence or absence of the drug for 30 min and then processed as described in Figure 1. Bars, 10 µm. (B) Total integrated fluorescence intensity of internalized Tfn and MHCI was quantified using Metamorph Software as described in Materials and Methods. The values of the different doses of pitstop 2 were then normalized to DMSO controls (set to 100%). Quantification was done for 60 cells at each dose and the error bars represent the standard deviation from the mean. P-values were calculated from the raw data and compared between DMSO and different doses of pitstop 2. Inhibition of MHCI uptake with pitstop was statistically significant for all doses with p-value <0.001 except for 5 µM pitstop 2 with p-value <0.02. Tfn uptake was also inhibited at 20 and 30 µM with p<0.001. The images and quantification shown are from one experiment; similar results were obtained in two additional experiments.