The Role of GATA1s in DS AMkL Figure 6. Down-regulation of GATA1s affects PF4, TUBB1, and IL1A expression in CMK cells. Panels A–C: Transcript levels for PF4 (panel A), TUBB1 (panel B), and IL1A (panel C) in the CMK-5a, -5b, and –neg cells were quantified by real-time RT-PCR as described in the Materials and Methods and results were expressed as mean values 6 standard errors from 3 independent experiments using the same cDNA preparation and normalized to GAPDH. Panel D: CMK-5a cells were incubated with or without 40 ng/mL recombinant IL1a protein for up to 48 h and counted in triplicate every 24 h. * indicates p,0.05. doi:10.1371/journal.pone.0027486.g006 Figure 7. IL1A is a bona fide GATA1s target gene in DS AMkL. In vivo binding of GATA1s to the putative GATA1 binding sites located in the upstream region of the IL1A gene (panel A) in CMK cells was determined by ChIP assays with use of regular PCR (panel B) and real-time PCR (panel C), as described in Materials and Methods. ** indicates p,0.005. doi:10.1371/journal.pone.0027486.g007 PLoS ONE | www.plosone.org 8 November 2011 | Volume 6 | Issue 11 | e27486