Male, female, and mixed groups under heat-shock (HS) and no-stress (n.S.) conditions. HSP70 was detected using a rat anti-HSP70 primary antibody and a donkey anti-rat secondary antibody with fluorescence detection at 700 nm (red). Actin was used as loading control and detected using a rabbit anti-actin primary antibody and a goat anti-rabbit secondary antibody at 800 nm (green). HSP70 signal intensity was quantified and normalized to the corresponding actin signal.