Figure S2, related to Figure 1and Figure 2. Genetic lesions in
pfk-1.1 mutants and endogenous expression pattern(A-B) Schematic of the genomic region of
pfk-1.1 and locations of the genetic lesions of the
pfk1.1 alleles examined in this study (marked with arrows) (A); and percentage of animals displaying a diffuse distribution of synaptic vesicle proteins in rescuing backgrounds (B). Single Nucleotide Polymorphism (SNP) mapping was performed and the
ola72 lesion was determined to be located between 2.77 Mb and 3.16 Mb on chromosome X (A). Fosmid WRM0616aB04, which contains the
pfk-1.1 genomic region (as well as un-named gene, Y41G9A.5), was shown to rescue the diffuse distribution of synaptic vesicle protein phenotype in
pfk-1.1(
ola72) mutant animals exposed to 10 minutes of hypoxic conditions (induced by mounting on glass slide with coverslip (Pitts and Toombs, 2004)) (B). The
pfk-1.1 promoter driving
pfk-1.1 cDNA also rescues the synaptic vesicle clustering phenotype in
pfk-1.1(
ola72) mutant animals exposed to 10 minutes of hypoxic conditions (B). Number of animals scored is indicated at the bottom of each column.
pfk-1.1(
ola72) is the same as shown in Figure 1J for (B).(C) Percentage of animals displaying a diffuse distribution of synaptic vesicle proteins after 10 minutes of hypoxic treatment (induced by mounting on glass slide with coverslip (Pitts and Toombs, 2004)). Note that all three independent
pfk-1.1 alleles phenocopy
pfk-1.1(
ola72), and that
pfk-1.1(
gk549413) fails to complement
pfk-1.1(
ola72). Number of animals scored is indicated at the bottom of each column. Wild-type control and
pfk-1.1 (
ola72) is the same as shown in Figure 1J. (D-G) Expression of the
pfk-1.1 promoter is observed in neurons of the ventral and dorsal nerve cords (arrows) (D); and in head neurons, including the NSM neurons (E-G).(H) Schematic of the glycolysis pathway and mutants examined in this study. Enzymes are shown in black, while substrates are in blue. Numbers inside parentheses indicate the number of identified C. elegans homologs for each given enzyme in the pathway. Solid red arrow denotes enzymatic reactions in the glycolysis pathway. Dashed arrows denote regulatory steps: (+) for activation and (-) for inhibition. Asterisk marks genes examined in this study. We selected the specific enzymes for analyses based on availability of identified alleles from the Caenorhabditis Genomic Center (CGC). With the exception of 6-phosphofructokinase/pfk-1.1, which was identified in our forward genetic screen, all other enzymes were selected based on theavailability of deletion alleles. Scale bar represents 25um for (D), and 5um for (E). Error bars denote SEM. *, p < 0.05. **, p <0.01. ***, p < 0.001 between indicated groups.