Fig. 4. Development-specific phosphorylation of p38 MAPK in nondiapausing flies at 0°C. Flesh flies were reared at a nondiapausing condition of (A) 25°C or (B) 20°C. Larvae or pupae were held at 0°C for 2 h, and protein samples were prepared. Fly proteins were analyzed using western blotting with anti-phospho-p38 and anti-total p38 MAPK antibodies. FL, 3^rd-instar feeding larvae; W0-2, day 0–2 wandering 3^rd-instar larvae; P0–12, day 0–12 after pupariation; NT, no treatment; 0°C, held at 0°C for 2 h. Red-eye stage nondiapausing pharate adults reared at 25°C were used as controls.