Temporal expression of HIF-independent hypoxic response genes.

<p>(A–H) Developmental hypoxic response profiles from qRT-PCR analyses are shown for transcripts that display HIF-independent (A – <i>LDH</i>, B – <i>phosphofructokinase</i>, C – <i>NMNAT</i>, D – <i>Alas</i>, E – <i>Cyp9b1</i>, F – <i>Cyp6a17</i>, G – <i>Cyp6a22</i>) or largely HIF-independent expression (H – <i>Peroxiredoxin 2540-2</i> – we note that <i>Prx2540-2</i> displays HD expression for probe set 1631628_s_at in the microarray and is thus classified as such; however, a second, non-overlapping probe set, 1633471_at, which is much more robustly induced in H appears to be from HI action, although its variability is too great to classify in this manner (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003230#pgen.1003230.s007" target="_blank">Table S1</a>). Our qPCR primer set favors HI expression.) Control (<i>w¹¹¹⁸</i>) animals or <i>sima</i> mutants were challenged for 6-hrs with 4% O₂. Developmental challenge times are identical to those from <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003230#pgen-1003230-g003" target="_blank">Figure 3</a>. All values are from experiments performed in triplicate from pools of biological replicates. Values are normalized to <i>rp49</i> expression and are reported as the relative fold-change of H/N. The dotted line represents no net change in response, or a value of 1.0. The vertical black line in late-L3 is when microarray analysis was performed. Error bars are the SEM. Separate normoxic and hypoxic traces for each transcript surveyed is shown in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003230#pgen.1003230.s004" target="_blank">Figure S4</a>.