Figure 6: Microarray analysis to identify GY118F downstream targets.

(a–c). Experimental samples are GY118F iPS cells derived and maintained in N2B27 plus G-CSF (0), same cells but from which G-CSF was withdrawn for 12 or 24 h (−12, −24) and that subsequently had G-CSF added back for 2 h and 40 min (−12+2h40m, −24+2h40m). vs, versus. (a) Genes found by microarray array analysis that were either downregulated on G-CSF withdrawal and upregulated on re-addition of G-CSF or upregulated on G-CSF withdrawal and downregulated on re-addition of G-CSF. Yellow represents gene expression downregulation and blue upregulation. Range from −4.3- to +4.3-fold change, with a minimum of 1.4-fold change. (b) Western blot analysis for Phospho-STAT3 and qRT–PCR analysis for known JAK/STAT3 targets Stat3, Socs3 and Klf4. Error bars indicate ±1s.d (n=3). (c) Western blot and q-RT–PCR analysis for pluripotency master regulators Oct4 and Nanog. Error bars indicate ±1 s.d. (n=3). (d) Genes that are upregulated in GY118F EpiSCs stimulated with G-CSF for 3 h, but not in controls, that were also found to be putative downstream targets of GY118F in iPS cells. (e) qRT–PCR analysis for Socs3, Stat3 and Klf4 in GY118F and control EpiSCs before (0), and after 3 h (3) of G-CSF stimulation. Error bars indicate ± 1 s.d (n=3).