Figure 1

FBXL19 induces Rac3 degradation - A. HEK293 cells were transfected with V5 or HA tagged FBXL19 (FBXL19-V5 or FBXL19-HA) plasmid for 48 h. Cells lysates were analyzed by immunoblotting with Rac3, V5 tag, HA tag, and β-actin antibodies. B. Rac3 levels in Figure 1A were quantified by Image J software. C. HEK293 cells were transfected with V5 tagged Nedd4L, FBXL18, or FBXL22 plasmid for 48 h. Cell lysates were analyzed by immunoblotting with Rac3, V5 tag, and β-actin antibodies. D. Cell lysates from MLE12, HEK293, A549, and OE19 were analyzed by immunoblotting with Rac3, Rac1, and β-actin antibodies. E. MLE12 cells were co-transfected with V5 tagged Rac3 (Rac3-V5) and FBXL19-V5 (0–2 μg) plasmids for 48 h and then the cell lysates were analyzed by immunoblotting with V5 tag and β-actin antibodies. F. MLE12 cells were co-transfected with Rac3-V5 and FBXL19-HA (0–4 μg) plasmids for 48 h and then the cell lysates were analyzed by immunoblotting with V5 tag, HA tag and β-actin antibodies. G. MLE12 cells were co-transfected with Rac3-V5, Cont shRNA, or one of the three FBXL19shRNAs (#1, #2, and #3) plasmids for 48 h and then the cell lysates were analyzed by immunoblotting with V5 tag, FBXL19 and β-actin antibodies. Rac3-V5 levels were quantified by Image J software. H. Total RNA was extracted from empty vector- or FBXL19-V5 plasmid-transfected HEK293 cells, Rac3 mRNA levels were then examined by RT-real time PCR with specifically designed Rac3 primers. All the blots are representative of three independent experiments.