Comparison of TIR1 transgene mediated degradation in C. elegans germline stem cells by qRT-PCR: (A) Schematic showing four distinct TIR1 transgenes driven by different promoters, indicated by the gene name followed by 'p', for promoter. mRuby and BFP are fluorescent proteins, F2A is a self-cleavable linker. Not shown is the substrate, LAG-1::degron. (B) Schematic illustrating sample collection for qRT-PCR analysis. The animals were dissected and the extruded gonads collected for RNA purification and then qRT-PCR analysis. (C & D) The relative mRNA level of LAG-1 dependent transcriptional targets
lst-1 (C) and
sygl-1 (D) from dissected gonad samples of the indicated genotype and treatment.
q175 is a
glp-1 null allele, where the mRNA level of
sygl-1 and
lst-1 were normalized by setting their levels to one for this genotype.
ar202 is a
glp-1 gain of function allele. Results are from three biological replicates, except for
sun-1p::TIR1 (two replicates). Bar shows mean and standard deviation. Each dot represents a single data point. ns for not significant, * for p<0.01, and ** for p<0.001 by two-tailed t-test.