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. 2016 May 18;6(5):160080. doi: 10.1098/rsob.160080

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© 2016 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

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Figure 3. — ChREBP and its effector lipogenic genes, ACC and FAS, are strongly upregulated in mature queens compared with kings and sterile individuals. (a) Expression of ChREBP mRNA in different castes and life stages of Prorhinotermes canalifrons. Total RNA was prepared and expression levels were determined by quantitative RT-PCR. Actin mRNA was used for normalization [29]. Reproductives were 4 months or 4 years old. Values are means ± s.e. (error bars) of at least 10 termites from each caste of four colonies reared in the laboratory. Significant differences (***p < 0.001) are indicated with asterisks. (b) Expression of ChREBP mRNA (qRT-PCR analysis) in different castes obtained during queenless experiments: in these conditions, some P. canalifrons workers are able to differentiate into male and female neotenics. Values are means ± s.e. (error bars) of three termites from three independent experiments. Significant differences from worker and male neotenics are indicated with asterisk (*p < 0.05). (c) Expression of ChREBP mRNA (qRT-PCR analysis) in female primary reproductives compared with workers in Prorhinotermes and in the Termitidae Aparatermes, Anoplotermes, Cavitermes, Neocapritermes, Embiratermes, Labiotermes and Nasutitermes. Columns represent the mean of collected workers (n = 10) and physogastric queens (n = 3). (d) Expression of ChREBP protein evaluated by western blot in P. canalifrons compared with mice tissues used as positive controls and efficiency of the antibodies (a representative blot is shown; upper panel: all proteins revealed by Coomassie staining; lower panel: western blot). Molecular marker (100 kDa) is provided. (1) Liver, (2) fat, (3) female reproductive (neotenic), (4) worker and (5) soldier. (e) Immunofluorescence image of the cellular ChREBP protein expression in a queen of P. canalifrons. Identification of nuclear (red arrow) and cytoplasmic (white arrow) ChREBP expression labelled using a commercial antibody against the human ChREBP peptide in green (left panel) and merged with nuclear DAPI stain in blue (right panel). Scale bars, 10 µm. (f) Expression of ChREBP, ACC and FAS mRNA (qRT-PCR analysis) in workers, male and female primary reproductives (king and queen, respectively), and female secondary reproductives (neotenic) of P. canalifrons. Values are means ± s.e. (error bars) of six termites from four independent colonies reared in the laboratory. Significant differences (**p < 0.01) from worker and male primary reproductive are indicated with asterisks. As values of workers are indicated as 1 arbitrarily for (a), (c) and (f), results are expressed according to the value of workers (fold).