Fig. 8. Analysis of Pol IIo^ser2 distribution and transcription at active loci during the heat shock response in JIL-1 mutant backgrounds. (A) Polytene chromosome squash preparations from JIL-1^z2/JIL-1^z2 (z2) and JIL-1^z2/JIL-1^z2 Su(var)3-9⁰⁶ (z2, 3-9) Drosophila larvae triple labeled with Pol IIo^ser2 antibody (green), Hsf antibody (red) and Hoechst (DNA, gray/blue) after heat shock treatment. Arrows point to heat shock puff regions. (B) Immunoblot of protein extracts from salivary glands from JIL-1^z2/JIL-1^z2 (z2) and JIL-1^z2/JIL-1^z2 Su(var)3-9⁰⁶ (z2, 3-9) larvae with (+HS) and without (-HS) heat shock treatment labeled with Pol IIo^ser2 antibody. Labeling with lamin antibody was used as a loading control. (C) Transcript levels of Hsp70 mRNA in JIL-1 null mutant backgrounds in response to heat shock treatment. Hsp70 transcript levels were determined by qRT-PCR and normalized to the mRNA levels of the control non-heat shock protein Rp49 (Ribosomal protein 49) both without and after heat shock treatment. The data shown are the average from two independent experiments in which total RNA was isolated from wild-type (wt), JIL-1^z2/JIL-1^z2 (z2) and JIL-1^z2/JIL-1^z2 Su(var)3-9⁰⁶ (z2, 3-9) larvae and each determination of transcript levels was performed in duplicate. Error bars indicate the s.d.m.