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. Author manuscript; available in PMC: 2012 May 13.
Published in final edited form as: Cell. 2011 May 13;145(4):596–606. doi: 10.1016/j.cell.2011.04.013

Figure 1. Reduced fat stores in SIK3 mutant flies, also see Figure S1.

Figure 1

(A) Diagram of SIK3 (CG15072) genomic locus is depicted. Location of P element EY06260 shown, along with SIK348 and SIK372 deletions generated by imprecise excision. SIK3+51 is a precise excision control line. SIK348 lacks the first (non-coding) exon, resulting in a significant reduction of SIK3 mRNA. The SIK372 deletion extends through the first two exons, removing sequences that encode part of the SIK3 kinase domain, and acts as a null allele.

(B) Q-PCR analysis of SIK3 mRNA levels in SIK3+ (SIK3+51) and SIK3− (SIK348) adult male and female flies under feeding conditions.

(C) Immunohistochemical analysis of SIK3 expression in fat bodies of WT (yw) and SIK3− (yw;SIK348/Df(2R)P34) L3 larvae. SIK3 protein shown in green, DNA shown in red. Scale bar; 10 μm.

(D, E) Relative fasting (24 hours) or ad libitum feeding lipid levels (D) and starvation sensitivities (E) of SIK3+ (SIK3+51) and SIK3− (SIK348) flies. (n=10 per genotype for lipid measurement and n=50 for starvation assays).

(F) Effect of fat body (FB) specific expression of SIK3 on lipid levels in SIK3− flies. FB>(FB-GAL4/+), SIK3−,FB>(FB-GAL4,SIK348/SIK348), SIK3−, FB>SIK3 (FB-GAL4, SIK348/SIK348;UAS-SIK3/+), SIK3−,UAS-SIK3(SIK348,UAS-SIK3/+) and FB>SIK3 (FB-GAL4/+; UAS-SIK3/+) flies are shown. (n=16 per genotype).

(G) Fat body specific expression of SIK3 rescues starvation sensitivity in SIK3− flies. Shown are survival curves for SIK3−, FB>SIK3 (FB-GAL4, SIK348/SIK348;UAS-SIK3/+), SIK3−, UAS-SIK3 (SIK348, UAS-SIK3/+) and SIK3−, FB> (FB-GAL4, SIK348/SIK348) flies. (n=97-127 per genotype).

(H) Lipid levels in SIK3 mutant flies with fat body specific expression of wild-type or catalytically inactive (K70M) SIK3, SIK2, and AMPK. Genotypes are: SIK3−, FB> (FB-GAL4, SIK348/SIK348), SIK3+, FB> (FB-GAL4/+), SIK3−, FB>SIK3.WT (FB-GAL4, SIK348/SIK348;UAS-SIK3/+), SIK3+, FB>SIK3.WT (FB-GAL4/+;UAS-SIK3/+), SIK3−, FB>SIK3.K70M (FB-GAL4, SIK348/SIK348;UAS-SIK3.K70M/+), SIK3+, FB>SIK3.K70M (FB-GAL4/+;UAS-SIK3.K70M/+), SIK3−, FB>SIK2 (FB-GAL4, SIK348/SIK348;UAS-SIK2/+), SIK3+, FB>SIK2 (FB-GAL4/+;UAS-SIK2/+), SIK3−, FB>AMPK (FB-GAL4, SIK348/SIK348;UAS-AMPKTD/+) and SIK3+, FB>AMPK (FB-GAL4/+;UAS-AMPKTD/+). (n=12 per genotype). Data are averages ± SEM (**, P<0.01; *, P<0.05).