(A-C) Dot plot of selected enriched GO-terms of early, consistent, and
late responder genes to sSpi over-expression, which are the
genes significantly changed only at 8h, only at 24h, or at both 8h and 24h (with
the same trend), respectively. For the full list of enriched GO-terms, see Data S4. Red indicates
significantly enriched terms for the up-regulated gene set, while green
indicates significantly enriched terms for the down-regulated gene set. The size
of the dot indicates the gene count for the GO-term, and the color indicates the
proportion of genes in the GO term that are significantly differentially
expressed. (D-E) Heatmap of (D) OXPHOS and (E) TCA cycle genes after
sSpi and Ets21C 24h over-expression. All
genes with known function involved in metabolic pathways (KEGG dme00190 and
dme00020) and expressed in ISCs were included. Black boxes on the right of the
heatmaps indicate DamID binding for the specified TF. Transcripts per million
(TPM) values were z-score normalized along rows before plotting. Genes were
manually ordered. (F) Gene Set Enrichment Analysis of sSpi 24h differentially
expressed genes showing OXPHOS, TCA, and Pyruvate Metabolism were significantly
up-regulated. (G) Venn diagram shows that Pnt and Ets21C differentially
expressed genes are essential for EGFR signaling early activation, making up to
56.35% of sSpi 8h transcription output. The overlap is
significant, P=1.24e-276. See also Figure S3.