Supplementary Materials

Supplementary Materials for:

PLC-γ and PI3K Link Cytokines to ERK Activation in Hematopoietic Cells with Normal and Oncogenic Kras

Ernesto Diaz-Flores, Hana Goldschmidt, Philippe Depeille, Victor Ng, Jon Akutagawa, Kimberly Krisman, Michael Crone, Michael R. Burgess, Olusegun Williams, Benjamin Houseman, Kevan Shokat, Deepak Sampath, Gideon Bollag, Jeroen P. Roose, Benjamin S. Braun, Kevin Shannon*

*Corresponding author. E-mail:

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  • Fig. S1. Characterization of the K+MG and M+G+ cell populations.
  • Fig. S2. Effects of various inhibitors on pERK and pTyr abundances in wild-type and Mx1-Cre, KrasG12D bone marrow cells.
  • Fig. S3. Abundance of signaling molecules in hematopoietic cells.
  • Fig. S4. Names and structures of the PI3K inhibitors used in this study.
  • Fig. S5. Effects of PI3K inhibitors on Ras-GTP abundance and on ERK and Akt phosphorylation in bone marrow cells stimulated with GM-CSF.
  • Fig. S6. Effects of PI3K inhibitors on pERK abundance in bone marrow M+G+ cells stimulated with GM-CSF.
  • Fig. S7. Effects of PI3K inhibitors on PLC-γ1 phosphorylation.
  • Fig. S8. JAK2, PLC-γ, and PI3K mediate GM-CSF–induced ERK phosphorylation in BMMPCs.
  • Fig. S9. Transduction efficiencies in BMMPCs and knockdown with shRNA constructs.
  • Fig. S10. PMA and GM-CSF induce Ras-GTP loading at different times in wild-type and KrasG12D BMMPCs.
  • Fig. S11. PMA-dependent activation of pERK is insensitive to inhibition of PI3K and PLC-γ, but not MEK.
  • Fig. S12. PLC-γ and CaMKII inhibitors, but not PKC inhibitors, block ERK phosphorylation.
  • Fig. S13. Basal amounts of phosphorylated PKC isoforms in wild-type BMMPCs and their response to GM-CSF.
  • Fig. S14. Effects of siRNA-mediated knockdown of RasGRP3 and RasGRP4 on ERK phosphorylation.
  • Fig. S15. Pharmacological activation of Raf largely overcomes the inhibitory effects of PI3K and PLC-γ inhibition.
  • Fig. S16. Dimethyl sulfoxide (DMSO) and U73343 do not alter pERK abundance in KLS CD48 cells.
  • Table S1. Sequences of PLC-γ1– and PLC-γ2–specific shRNAs.

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Citation: E. Diaz-Flores, H. Goldschmidt, P. Depeille, V. Ng, J. Akutagawa, K. Krisman, M. Crone, M. R. Burgess, O. Williams, B. Houseman, K. Shokat, D. Sampath, G. Bollag, J. P. Roose, B. S. Braun, K. Shannon, PLC-γ and PI3K Link Cytokines to ERK Activation in Hematopoietic Cells with Normal and Oncogenic Kras. Sci. Signal. 6, ra105 (2013).

© 2013 American Association for the Advancement of Science