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. Author manuscript; available in PMC: 2013 Nov 13.
Published in final edited form as: Cancer Cell. 2012 Nov 13;22(5):656–667. doi: 10.1016/j.ccr.2012.08.027

Figure 5. Inhibition of pre-BCR or ROR1 result in counter-balancing effects on signaling.

Figure 5

(A) RCH-ACV cells were treated with dasatinib for 24 hours or ROR1 siRNA for 72 hours and whole cell extracts were analyzed by phospho-proteomic array. Values on the graph represent percent phosphorylation change ± s.e.m. (n = 3) (*p<.05) of a total of 46 phospho-proteins for dasatinib relative to cells in the absence of any drug and for ROR1 siRNA relative to cells transfected with non-specific siRNA.

(B) RCH-ACV cells were transfected with ROR1 or non-specific siRNA for 72 hours and whole cell extracts were subjected to immunoblot analysis for ROR1, phospho-AKT at residue Serine 473 and β-Actin; ROR1, total and phospho-ERK, and β-Actin; ROR1, total and phospho-MEK, and β-Actin for validation of ROR1 modulation observed by the phospho-proteomics screen in (A).

(C) RCH-ACV cells were treated with dasatinib (50nM), PD98059 (50µM), or both over a time course and whole cell extracts were subjected to immunoblot analysis for total or phospho-AKT at residue Serine 473, ROR1, and β-Actin.

(D) RCH-ACV cells were electroporated in the presence of non-specific, ROR1-targeting, or Igα-targeting siRNA and then plated in culture media. After 2 days, graded concentrations of dasatinib were added. Cells were allowed to culture an additional 2 days before they were subjected to an MTS assay for measurement of cell viability. Values represent percent mean (normalized to no-drug control wells) ± s.e.m. (n = 5). (*p<.05, **p<.01). See also Figure S6.