Figure 4. GFP-PODXL mobility is increased by PDZ-binding motif deletion or NHERF1 knockdown. (A) FRAP measurements on cells 4 d after seeding show that the lateral mobility of GFP-PODXL was increased either by deletion of the last four amino acids of its cytoplasmic tail (GFP-PODXLΔ4) or by knockdown of NHERF1 (NHERF1 shRNA). GFP-GPI is thought to freely diffuse in the membrane and recovers faster than the GFP-PODXL constructs. (B) FRAP measurements taken 120 s after photobleaching show a significant difference in FRAP between GFP-PODXL and GFP-PODXLΔ4 (P < 0.005). A similar difference was found between FRAP of GFP-PODXL after shRNA knockdown of NHERF1 and vector controls (P < 0.005). Values are means of FRAP measurements from 10 cells for GFP-GPI, GFP-PODXL, GFP-PODXLΔ4, and GFP-GPI and 8 cells for vector and NHERF1 shRNA. Error bars represent standard deviation between measurements.
Figure 4.

GFP-PODXL mobility is increased by PDZ-binding motif deletion or NHERF1 knockdown. (A) FRAP measurements on cells 4 d after seeding show that the lateral mobility of GFP-PODXL was increased either by deletion of the last four amino acids of its cytoplasmic tail (GFP-PODXLΔ4) or by knockdown of NHERF1 (NHERF1 shRNA). GFP-GPI is thought to freely diffuse in the membrane and recovers faster than the GFP-PODXL constructs. (B) FRAP measurements taken 120 s after photobleaching show a significant difference in FRAP between GFP-PODXL and GFP-PODXLΔ4 (P < 0.005). A similar difference was found between FRAP of GFP-PODXL after shRNA knockdown of NHERF1 and vector controls (P < 0.005). Values are means of FRAP measurements from 10 cells for GFP-GPI, GFP-PODXL, GFP-PODXLΔ4, and GFP-GPI and 8 cells for vector and NHERF1 shRNA. Error bars represent standard deviation between measurements.

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