Figure S9 Expression pattern of
aakb-1. A) Confocal images showing Paakb-1::GFP in 1-day old wild type hermaphrodites. The Paakb-1::gfp reporter contains 2.845 Kb of promoter directly upstream of the transcriptional start site. (i) Whole worm expression pattern. (ii) Paakb-1::GFP is strongly expressed in pharyngeal muscles (PhxM), the pharyngeal intestinal valve cells (VPI) as previously noted [51] and a few head neurons of the pharynx (PhxN) and the ring region (RingN). It is also detected in some support cells (SC). (iii) Tail expression of Paakb-1::GFP includes the tail minor epithelium (TE), the rectal-intestinal valve (RectV), posterior intestine and body wall muscles (BWM). (iv) Mid-body expression is mostly restricted to muscles of the vulva (VlvM), uterus (UtM) and body wall (BMW), but is also detected in the intestine (Int). However we noted that the levels of GFP expression in all genetic backgrounds was highly variable between animals. Table S10 compares expression of Paakb-1::gfp with other AMPK subunits. B) Quantification of GFP fluorescence in the heads of worms expressing a Paakb-1::GFP reporter. The
daf-16;
daf-2 background carrying this transgene exhibited unexplained and higher than normal intestinal autofluorescence making fluorescence quantification of whole worms difficult. To circumvent this we measured GFP fluorescence levels in the heads of worms, where Paakb-1::GFP is highly expressed. Mean of four trials shown. Error bars, standard deviation. C) qRT-PCR of gfp mRNA in worms expressing a Paakb-1::GFP reporter revealed an increase in expression between
daf-2 and
daf-2;
daf-16, however this was not significant, probably due to transgene expression varying between individuals. Error bars, standard deviation.