Figure S1.
Leep2 complex translocates to the cell periphery in response to chemoattractant stimulation. (A) Schematic illustration of the domain organization of Leep2 and human RalGAP complexes. (B) Phylogenetic tree of Leep2 and human RalGAP complexes. (C) Quantification of the fluorescent intensity of GFP-Leep2A at macropinocytic cups in WT cells, with or without the expression of RFP-Leep2B. The scatter plot shows data points with means and SD (n represents the number of cells analyzed). Significance was determined by two-tailed unpaired t test. (D) Translocation of GFP-Leep2A and RFP-Leep2B in response to cAMP stimulation (1 μM cAMP was added at time 0). Top: Time-lapse imaging of translocation. Bottom: Quantification of translocation (mean ± SD, n represents the number of cells analyzed). (E) Translocation of GFP-Leep2A and RFP-Leep2B in response to folic acid (FA) stimulation (50 μM FA was added at time 0). Top: Time-lapse imaging of translocation. Bottom: Quantification of translocation (mean ± SD, n represents the number of cells analyzed). (F) Translocation of GFP-Leep2A and RFP-Leep2B in response to cAMP stimulation (1 μM cAMP was added at time 0) in the presence of Latrunculin A (LatA). Top: Time-lapse imaging of translocation. Bottom: Quantification of translocation (mean ± SD, n represents the number of cells analyzed). Scale bars, 5 μm.

Leep2 complex translocates to the cell periphery in response to chemoattractant stimulation. (A) Schematic illustration of the domain organization of Leep2 and human RalGAP complexes. (B) Phylogenetic tree of Leep2 and human RalGAP complexes. (C) Quantification of the fluorescent intensity of GFP-Leep2A at macropinocytic cups in WT cells, with or without the expression of RFP-Leep2B. The scatter plot shows data points with means and SD (n represents the number of cells analyzed). Significance was determined by two-tailed unpaired t test. (D) Translocation of GFP-Leep2A and RFP-Leep2B in response to cAMP stimulation (1 μM cAMP was added at time 0). Top: Time-lapse imaging of translocation. Bottom: Quantification of translocation (mean ± SD, n represents the number of cells analyzed). (E) Translocation of GFP-Leep2A and RFP-Leep2B in response to folic acid (FA) stimulation (50 μM FA was added at time 0). Top: Time-lapse imaging of translocation. Bottom: Quantification of translocation (mean ± SD, n represents the number of cells analyzed). (F) Translocation of GFP-Leep2A and RFP-Leep2B in response to cAMP stimulation (1 μM cAMP was added at time 0) in the presence of Latrunculin A (LatA). Top: Time-lapse imaging of translocation. Bottom: Quantification of translocation (mean ± SD, n represents the number of cells analyzed). Scale bars, 5 μm.

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