Fig. 2

BMP signaling suppresses AMPK and AMPK negative feedback regulation of BMP in C. elegans. A Western blot analysis for pAMPK of C. elegans with BMP loss-of function (lof) mutants of BMPR2 daf4(e1364) and Smad transcription factors (sma-6 and sma-2) compared to wild-type worms. AMPK lof mutants aak-2 (ok524) were utilized as a control. Nematodes were grown at 20 °C until L4 stage. Each lane represents 25 animals. B Fluorescence images and quantification of C. elegans expressing the AMPK downstream reporter nhr-49p::nhr-49::GFP with genotypes harboring BMP lof (SMA-6) and gof lon-1( e185),lon-2 ( e678) (n = 30 animals). Images taken at L4 stage animals under well-fed conditions at 20 °C. Scale bar indicates 50 microns. C Fluorescence quantification of C. elegans expression of the BMP transcriptional reporter spp-9::GFP in AMPK wild-type and AMPK lof mutant aak-2 (ok524) in either well-fed animals or after 8 h of starvation (n = 30 animals). D Body-length measurements of indicated genotypes at L4 stage grown at 20 °C (n = 30 animals). E Relative quantification of mRNA levels of indicated genes in wild-type and AMPK gof mutant aak-2 animals at L4 stage grown at 20 °C. All alleles are loss of function except for aak-2(gof), which is a transgene that expresses a truncated, constitutively active kinase