Supplemental Tables for "Hypoxia-inducible factor 1alpha is required to establish the larval glycolytic program in <i>Drosophila melanogaster</i>"

<b>Objectives:</b> The rapid growth that occurs during <i>Drosophila</i> larval development requires a dramatic rewiring of central carbon metabolism to support biosynthesis. Larvae achieve this metabolic state, in part, by coordinately up-regulating the expression of genes involved in carbohydrate metabolism. The resulting metabolic program exhibits hallmark characteristics of aerobic glycolysis and establishes a physiological state that supports growth. To date, the only factor known to activate the larval glycolytic program is the <i>Drosophila</i><i> </i>Estrogen-Related Receptor (dERR). However, dERR is dynamically regulated during the onset of this metabolic switch, indicating that other factors must be involved. Here we examine the possibility the <i>Drosophila</i><i> </i>ortholog of Hypoxia inducible factor 1a (Hif1α) is also required to activate the larval glycolytic program.<b>Methods:</b><b> </b>CRISPR/Cas9 was used to generate new loss-of-function alleles in the <i>Drosophila</i><i> </i>gene <i>similar</i><i> </i>(<i>sima</i>), which encodes the sole fly ortholog of Hif1a. The resulting mutant strains were analyzed using a combination of metabolomics and RNAseq for defects in carbohydrate metabolism.<b>Results:</b> Our studies reveal that <i>sima</i><i> </i>mutants fail to activate aerobic glycolysis and die during larval development with metabolic phenotypes that mimic those displayed by <i>dERR</i><i> </i>mutants. Moreover, we demonstrate that dERR and Sima/Hif1α protein accumulation is mutually dependent, as loss of either transcription factor results in decreased abundance the other protein.<b>Conclusions:</b><b> </b>These findings demonstrate that Sima/HIF1a is required during embryogenesis to coordinately up-regulate carbohydrate metabolism in preparation for larval growth. Notably, our study also reveals that the Sima-dependent gene expression profile shares considerable overlap with that observed in <i>dERR</i><i> </i>mutant, suggesting that Sima/HIF1a and dERR cooperatively regulate embryonic and larval glycolytic gene expression.