An intersectional gene regulatory strategy defines subclass diversity of C. elegans motor neurons

*Reagent: wdIs3 X or wdIs6 II [del-1^prom::gfp]; Winnier et al., 1999.

^†Expression was confirmed in VA1-11, VB1-11, and SABVL/R based on cell body position, axonal trajectory, and expected number of neurons for each class. Also, see Figure 2C.

^‡As described in Winnier at al. (1999), by the end of L2, del-1^prom::gfp is visible in a few anterior VAs. This expression progresses in a wave from anterior to posterior VAs, with all VAs (except VA12) expressing del-1^prom::gfp by the L4/adult stage.

^§Reagent: evIs82B IV [unc-129^prom::gfp]; Colavita et al. (1998), Kratsios et al. (2015)

^#Expression was confirmed in DA1-7, DB1-7, and SABD based on cell body position, axonal trajectory, cell type-specific markers, and expected number of neurons for each class. Also, see Figure 2C.

^¶Reagent: mdIs123 [hum-2^prom::gfp] from James Rand and Stephen Fields; www.wormatlas.org

**Expression was confirmed in DB1 based on cell body position and axonal trajectory. Also, see Figure 1B.

^††Reagent: bnc-1 (ot845[bnc-1::+mNG+AID]) CRISPR allele; Kerk et al., 2017.

^‡‡Expression was confirmed in VA1-VA12, VB2-VB11, SABVL/R based on cell body position and expected number of neurons for each class.

^§§Reagent: otEx6502 [cfi-1^fosmid::gfp]; Kerk et al., 2017.

^##Expression was confirmed in DA1-8 based on cell body position and the cell-type specific markers juIs14[acr-2::gfp] that labels DA, DB, VA, VB. This analysis also revealed that cfi-1 is not expressed in VA12. No expression in VA1, VB1 and VB2 was inferred based on cell body position and total number (3) of MNs expressing cfi-1 at the retrovesicular ganglion.

^¶¶Reagent: adEx1299[avr-15^prom::gfp]; Dent et al., 1997.

***Expression was confirmed in AS11 based on cell body position, axonal trajectory, and cell type-specific markers. Also, see Figure 9D–E and Figure 1-figure supplement 1 .

^†††Reagent: ynIs59[flp-18^prom::gfp] from Chris Li. Expression in VA11 and VA12 was confirmed with cell type-specific markers. Also, see Figure 1B, Figure 2A–B, and Figure 1—figure supplement 1.

^‡‡‡Reagent: otIs107 [ser-2^prom1::gfp]; Tsalik et al., 2003.

^§§§Expression in DA8 was confirmed with cell type-specific markers. Also, see Figure 1B and Figure 1—figure supplement 1.

^###Reagent: muIs42[mig-13^prom::MIG-13::gfp]; Sym et al., 1999, Klassen and Shen (2007)

^¶¶¶Expression in DA9 and VA12 was confirmed with cell type-specific markers. Also, see Figure 2A–B.

****Reagent: otIs476[glr-4^prom::tagrfp]. Expression in DA9 was confirmed with cell type-specific markers. Also, see Figure 1—figure supplement 1.

^††††Reagent: otIs453 [itr-1^prom::gfp]. Plasmid used for otIs453 transgene is pBT001, which was generated by Gower et al., 2001 and gfp expression in C.elegans in DA9 and PDA is described in Gower et al., 2001 and Klassen and Shen (2007). Expression was confirmed in DA9 using cell type-specific markers. Also, see Figure 2A–B and Figure 1—figure supplement 1.

^‡‡‡‡Reagent: sEx11976[sra-36^prom::gfp]. Expression in VA11, DA8 and DA9 was confirmed with cell type-specific markers. Also, see Figure 1B and Figure 1—figure supplement 1. ^{†, #,} **^{, ‡‡, ##, ¶¶, †††, §§§, ###,} ****^{, ††††}See also ‘Motor Neuron Subclass Identification’ section in Materials and methods.

N. D: Not determined.