Exogenous SHP2 rescues C/EBPα expression and granulopoiesis after SHP2 knockdown. (A) 32Dcl3(shRNA77-10) cells were transduced with pBabeNeo (Neo) or pBabeNeo expressing the SHP2 cDNA modified to resist targeting the shRNA-77 without altering the encoded protein sequence (SHP2). These cells and 32Dcl3 cells transduced with the empty pLKO.1 vector (Puro) were cultured in IL-3 or in G-CSF for 1 day (G1). Total cellular proteins were collected and subjected to Western blotting for SHP2, C/EBPα, and β-actin. (B) Total cellular RNAs prepared from 32Dcl3(shRNA-77)-10 cells transduced with exogenous SHP2 or the empty Neo vector were prepared 1 day after transfer to G-CSF and analyzed for relative expression of MPO, PR3, NE, and G-CSFR (GR) RNAs. The mean relative expression of each mRNA after SHP2 rescue was set to 1.0. Results shown are mean and SE of 3 repetitions. * indicates P < .05 for each RNA after SHP2 rescue compared with mock rescue. (C) 32Dcl3-Puro, 32Dcl3(shRNA-77)-10, or 32Dcl3(shRNA-77)-10+SHP2 cells cultured in G-CSF for 5 days were cytospun and subjected to Wright-Giemsa staining.