Figure 2

Q-RT-PCR verification of selected microarray results and transcriptional analysis of Mk genes. (A) For transcriptional analysis beginning on day 5, Mk culture samples were enriched for CD41a⁺ cells by positive immunomagnetic selection. Flow cytometric analysis after selection revealed high purity. Data is shown for one representative sample point from one culture. Open line: isotype control; Filled line: cells after selection. (B) Q-RT-PCR validation of microarray results across multiple Mk culture samples. Microarray log expression ratios from Mk culture samples are compared to values obtained using Q-RT-PCR for each of 3 genes – BBC3 (squares), MAP3K5 (diamonds), and SIRT7 (triangles). (C) Expression profiles for 33 Mk genes that were differentially expressed with time in Mk cultures and/or between Mk and G cultures. Color denotes degree of differential expression (saturated red = 4-fold up-regulation, saturated green = 4-fold down-regulation, blank = unchanged, gray = no data available). The first block shows average expression ratios across the biological replicates (n = 3) for the designated samples as compared to day 0 CD34⁺ cells; the second block shows expression ratios, averaged across the biological replicates (n = 3), as compared to the average expression from days 1 – 4; the third block shows average expression profiles as compared to equivalent-day G cells (day 12 Mk vs. day 11 for G cells) (n = 2); and the last block shows expression profiles of G cells with respect to average expression of G cells on days 1–4 (n = 2). The median ratio, along with the maximum (for up-regulated genes) or minimum (for down-regulated genes) ratio of Mk cells on days 5 – 12 (with respect to average expression of days 1–4) is provided (a negative value represents down-regulation).