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. 2017 Sep 15;144(18):3232–3240. doi: 10.1242/dev.151134

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© 2017. Published by The Company of Biologists Ltd

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Fig. 5. — Neither JIL-1 nor H2Av is required for chromatin decondensation or transcriptional elongation during heat shock. (A) 87 A/C heat shock puffs from polytene squash preparations labeled with Pol II0^ser2 and Pol II0^ser5 antibodies (green) from wild-type, homozygous H2Av⁸¹⁰ null, homozygous JIL-1^z2 null (z2), and homozygous JIL-1^z2 null larvae expressing the CFP-tagged JIL-1-CTD transgene (JIL-1-CTD; z2). DNA (Hoechst) is in blue. (B) Normalized 87 A/C puff size from wild-type, homozygous H2Av⁸¹⁰ null, homozygous JIL-1^z2 null (z2), and homozygous JIL-1^z2 null larvae expressing the CFP-tagged JIL-1-CTD transgene (JIL-1-CTD; z2). Measurements were obtained from more than 30 salivary gland nuclei from at least five different larvae for each genotype. The box plot representation defines 25th to 75th percentiles (boxes), 50th percentile (lines in boxes), ranges (whiskers, 1.5 times the interquartile range extended from both ends of the box or the maximal/minimal value), and outliers more than 3/2 times the upper quartile (circles). There was no statistically significant difference in puff size between the four genotypes (P>0.15; ANOVA test). (C) The distribution of salivary gland nuclei with clearly recognizable puffs among the total number of nuclei examined from wild-type, homozygous H2Av⁸¹⁰ null, homozygous JIL-1^z2 null (z2), and homozygous JIL-1^z2 null larvae expressing the CFP-tagged JIL-1-CTD transgene (JIL-1-CTD; z2). The total number of nuclei examined is indicated for each genotype. (D) Immunoblots of protein extracts labeled with Pol II0^ser2 and Pol II0^ser5 antibody from wild-type, homozygous H2Av⁸¹⁰ null, homozygous JIL-1^z2 null (z2), and homozygous JIL-1^z2 null larvae expressing the CFP-tagged JIL-1-CTD transgene (JIL-1-CTD; z2). Labeling with lamin antibody provided a loading control. (E) Transcript levels of Hsp70 mRNA in wild-type and homozygous H2Av⁸¹⁰ and JIL-1^z2 null mutant backgrounds in response to heat shock treatment. Hsp70 transcript levels were determined by qRT-PCR and normalized to the mRNA levels for the control non-heat shock protein Rp49 both without and after heat shock treatment. The data shown are the average from three independent experiments in which each determination of transcript levels was performed in duplicate. The error bars indicate s.d.