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. 2011 Apr 7;7(4):e1001331. doi: 10.1371/journal.ppat.1001331

Figure 5. THP-1 cells stably expressing K5 WT have sunitinib-sensitive increased RTK phosphorylation.

Figure 5

(A) Normalized whole cell lysates from THP-1 cells, mock (PBS) or sunitinib (2 µM) treated, were subjected to western blot using an anti-phospho-tyrosine (4G10) (pTyr) antibody (top panel) and reprobed with an anti-actin antibody (bottom panel) to demonstrate equal loading. (B) Normalized WCL from parental or K5 WT-expressing THP-1 cells were subject to immunoprecipitation (IP) using anti-Flt-3 antibodies followed by western blot (WB) using an anti-phospho-tyrosine (4G10) (pTyr) antibody (top panel). The IP (middle panel)and whole cell lysates (WCL) (bottom panel) were probed with anti-Flt-3 antibodies, as controls. (C) 293T cells were co-transfected with expression constructs for K5 and Flt-4 or a Flt-4 mutant (G857R). Two days post-transfection, cell lysates were subjected to IP using anti-Flt-4 antibodies. western blot (WB) was performed using anti-phospho-tyrosine (4G10) (pTyr) antibodies (top panel) and re-probed with anti-Flt-4 antibodies (bottom panel). Data for all panels are representative of three independent experiments.