Figure 8.
Leep2 complex regulates macropinocytosis through RasB, RasD, and RasG. (A) TRITC-Dextran uptake in WT and the indicated ras mutants. Independent clones of rasD and rasG double knockout cells were generated by deleting rasG in a rasD knockout background (rasD−rasG−) or deleting rasD in a rasG knockout background (rasG−rasD−). (B) Quantification of TRITC-Dextran uptake, as shown in A. (C) TRITC-Dextran uptake in WT cells expressing constitutively active RasB, RasD, or RasG under a doxycycline-inducible promoter. (D) Quantification of TRITC-Dextran uptake, as shown in C. (E) Scatter plots showing a negative correlation between fluorescence intensity (I) corresponding to GFP-fusion proteins and TRITC-dextran. The correlation coefficients are shown on the right. (F) Localization of Byr2RBD and Leep2B in WT cells co-expressing Byr2RBD-GFP, Flag-Leep2A, and RFP-Leep2B. (G) Quantification of the changes in fluorescent intensity of RFP-Leep2B and Byr2RBD-GFP on newly formed macropinosomes, as shown in F. The frame at which the macropinocytic cup closed was set as time 0, and the fluorescence intensity at other time points was normalized to that at time 0. (H) Localization of Byr2RBD-GFP in WT and DKO cells during macropinocytosis. (I) Quantification of the fluorescent intensity of Byr2RBD-GFP at the macropinocytic cups in WT and DKO cells. (J) Quantification of the changes in fluorescent intensity of Byr2RBD-GFP on newly formed macropinosomes. The frame at which the cup closed was set as time 0. The scatter plots in B, D, and I show data points with means and SD; n represents the number of cells analyzed. The plots in G and J show means and SD; n represents the number of cells analyzed. Significance was determined by one-way ANOVA in B and D and by two-tailed unpaired t test in I. Scale bars, 5 μm.

Leep2 complex regulates macropinocytosis through RasB, RasD, and RasG. (A) TRITC-Dextran uptake in WT and the indicated ras mutants. Independent clones of rasD and rasG double knockout cells were generated by deleting rasG in a rasD knockout background (rasD−rasG−) or deleting rasD in a rasG knockout background (rasG−rasD−). (B) Quantification of TRITC-Dextran uptake, as shown in A. (C) TRITC-Dextran uptake in WT cells expressing constitutively active RasB, RasD, or RasG under a doxycycline-inducible promoter. (D) Quantification of TRITC-Dextran uptake, as shown in C. (E) Scatter plots showing a negative correlation between fluorescence intensity (I) corresponding to GFP-fusion proteins and TRITC-dextran. The correlation coefficients are shown on the right. (F) Localization of Byr2RBD and Leep2B in WT cells co-expressing Byr2RBD-GFP, Flag-Leep2A, and RFP-Leep2B. (G) Quantification of the changes in fluorescent intensity of RFP-Leep2B and Byr2RBD-GFP on newly formed macropinosomes, as shown in F. The frame at which the macropinocytic cup closed was set as time 0, and the fluorescence intensity at other time points was normalized to that at time 0. (H) Localization of Byr2RBD-GFP in WT and DKO cells during macropinocytosis. (I) Quantification of the fluorescent intensity of Byr2RBD-GFP at the macropinocytic cups in WT and DKO cells. (J) Quantification of the changes in fluorescent intensity of Byr2RBD-GFP on newly formed macropinosomes. The frame at which the cup closed was set as time 0. The scatter plots in B, D, and I show data points with means and SD; n represents the number of cells analyzed. The plots in G and J show means and SD; n represents the number of cells analyzed. Significance was determined by one-way ANOVA in B and D and by two-tailed unpaired t test in I. Scale bars, 5 μm.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal