Figure S1 (related to Fig.2,3): Additional characterization of
bnc-1 and
mab-9A: Quantification for Fig.2F; neurons in the RVG where this
unc-3 promoter expresses poorlywere strictly excluded; error bars show SD. Identity of remaining VNC MNs where
unc129 was not repressed by the
unc-3prom::
bnc-1 transgene could not be unambiguouslyascertained as their positions seemed random and were thus binned together as D/V.Three independent transgenic lines were assessed (#1.1, #3.1, and #5.1). Unpaired ttests were performed comparing each line with its corresponding no-transgene controland WT; ***p < 0.001; n 10.B: The expression of VA/VB-specific
del-1 and
inx-12, DB/VB-specific
acr-5, and cholinergicunc-17 is not affected in
bnc-1 null mutants. Error bars show SD; n = 13.C: The axonal morphology of VA/VB (which project ventrally) and DA/DB MNs (which projectdorsally) is unaffected in, respectively,
bnc-1 and
mab-9 null mutants when compared toWT worms. The fluorophores of the reporter transgenes fill up the neuronal processes,allowing the axonal projections to be visualized. n=20.D: The distribution of presynaptic puncta of VA/VB (along the VNC) and DA/DB MNs (alongthe dorsal nerve cord) is unaffected in, respectively,
bnc-1 and
mab-9 null mutants whencompared to WT worms. RAB-3 (a member of the Ras GTPase superfamily) is apresynaptic molecule which when translationally fused with a fluorophore, allows for thevisualization of the presynapse of a neuron in which it is expressed. n=20.E: In C. elegans, cholinergic MNs in the VNC are dyadic, as they synapse onto muscle aswell as GABAergic MNs. The distribution of postsynaptic puncta in GABAergic MNs ofsynapses originating from VA/VB (along the VNC) and DA/DB MNs (along the dorsalnerve cord) is unaffected in, respectively,
bnc-1 and
mab-9 null mutants when comparedto WT worms. ACR-12 (an nAChR subunit) is a postsynaptic molecule which whentranslationally fused with a fluorophore, allows for the visualization of the postsynapse ofthe neuron in which it is expressed. In this case, the
unc-47 promoter drives ACR-12expression in GABAergic MNs. All worm strains here are in the genetic background ofacr-12
(ok367) which is required for ACR-12+GFP to be detectable. n = 20.F: The distribution of postsynaptic puncta in neuromuscular junctions (NMJ) originating fromVA/VB (along the VNC) and DA/DB MNs (along the dorsal nerve cord) is unaffected in,respectively,
bnc-1 and
mab-9 null mutants when compared to WT worms. Here, the endogenous
unc-29 locus (encoding an nAChR subunit that is a body wall muscle-specificpostsynaptic molecule) is translationally fused with TagRFP to allow for the visualizationof NMJ postsynapses. n = 20.G: VA/VB-specific expression of the
bnc-1 translationally tagged fosmid (very dim butconsistent) and transcriptionally tagged rescue construct reporters (as mentioned inFig.3E).H: Quantification for Fig.3E; error bars show SD. Unpaired t-tests were performed comparedto WT; ***p < 0.001; n 13.I: Images of the
unc-3-dependent, VA/VB-specific
lgc-36 (Gendrel et al., 2016) beingderepressed in DA/DB MNs in
mab-9 null mutants as quantified in Fig.3G. Showinganterior half of worm.J: The expression of DB/VB-specific
acr-5 is not affected in
mab-9 null mutants. Error barsshow SD; n=13.K: Quantification for
unc-3prom::
mab-9 transgenic line effect in Fig.3A,F; neurons in the RVGwhere this
unc-3 promoter expresses poorly were strictly excluded; error bars show SD.Identity of remaining VNC MNs where
del-1 was not repressed by the transgene could notbe unambiguously ascertained as their positions seemed random and were thus binnedtogether as D/V. At least two independent transgenic lines were assessed although onlythe most representative line is shown due to space constraints. Unpaired t-tests wereperformed comparing each line with its corresponding no-transgene control and WT; ***p< 0.001; n=10.