Figure 3.

Combination of HT with RT affects the main signaling networks and stress response pathways in HNSCC spheroids: (a) Venn diagrams illustrating the number of differentially expressed genes from the whole genome for RT, HT, and HT + RT-treated FaDu and SAS spheroids in triplicate (0.5 h after treatment). Genes were selected by DESeq2 with at least 2-fold changes in expression relative to the appropriate control spheroids (adjusted p ≤ 0.05). (b) The top 20 selected signaling pathways and processes that are significantly (2-fold change or more, adjusted p ≤ 0.05) over-represented in FaDu and SAS spheroids 0.5 h after exposure to HT + RT (42.5 °C/60 min; 7 Gy for FaDu and 10 Gy for SAS) when compared to controls using the GO enrichment analysis. The X-axis presents the corresponding adjusted p values according to Fischer exact’s test (in negative log10 scale). (c) Representative Western blot data sets showing the expression/activation of proteins of interest from the MAPK and PKB/Akt signaling pathways in the two HNSCC spheroid types upon treatment. GAPDH was used as loading control. Spheroid treatment conditions: Ctrl—control; RT—single dose irradiation (7 Gy for FaDu and 10 Gy for SAS); HT—hyperthermia (42.5 ˚C/60 min); HT + RT—hyperthermia and single dose irradiation according to mono-treatments.