Skip to main content
. Author manuscript; available in PMC: 2023 Apr 25.
Published in final edited form as: Curr Biol. 2022 Mar 7;32(8):1715–1727.e8. doi: 10.1016/j.cub.2022.02.040

Figure 6. CUT genes cooperate with terminal selectors to control pan-neuronal gene expression.

Figure 6.

(A) Illustration for how terminal selectors contribute to the regulation of pan-neuronal gene expression.

(B-H) Expression of ric-4(syb2878[ric-4::GFP]) (ric-4(syb2878[ric-4::T2A-3xNLS-GFP])) in wild-type (top left), terminal selector mutant (unc-86(ot1184) (B-F), ceh-14(ot1185) (G) or unc-30(ot1186) (H); top right), CUT sextuple mutant (bottom left), and compound terminal selector and CUT sextuple mutant (bottom right). Lateral views of the head (B), midbody (C-E, H) and tail (F and G) are shown. All images correspond to worms at the L4 larval stage, except for HSN (E) where young adults are shown. Quantification of ric-4(syb2878[ric-4::T2A-3xNLS-GFP]) fluorescence intensity in individual neurons. The data are presented as individual values with each dot representing the expression level of NSM (B), BDU (C), ALM (D), HSN (E), PLM (F), PHA, PHB, PVC (G), DD4, or VD8 (H) neuron, with the mean ± SEM indicated. Wild-type data is represented with black dots, terminal selector mutants with green dots, the sextuple CUT mutant with purple dots, and compound terminal selector and CUT sextuple mutant with yellow dots. One-way ANOVA followed by Tukey’s multiple comparisons test; *P < 0.05, **P < 0.01, ***P < 0.001. n ≥ 8 for all genotypes.

(I) Expression of ric-4(syb2878[ric-4::T2A-3xNLS-GFP]) in wild-type (top), CUT sextuple mutant (middle), and upon mutation of HOX and terminal selector binding sites on the ric-4 endogenous locus in a CUT sextuple mutant background (bottom). Individual mutation of the HOX (ric-4(ot1182 syb2878)) or terminal selector binding sites (ric-4(ot1181 syb2878)) has no effect on ric-4(syb2878[ric-4::T2A-3xNLS-GFP]) expression, but the expression is reduced in posterior ventral nerve cord (VNC) neurons when binding site mutations are combined (ric-4(ot1183 ot1181 syb2878)). Lateral views of the posterior VNC in L4 worms are shown. Quantification of ric-4(syb2878[ric-4::T2A-3xNLS-GFP]) fluorescence intensity in posterior VNC neurons. The data are presented as individual values with each dot representing the expression level of one worm with the mean ± SEM indicated. Wild-type data is represented with black dots, the sextuple CUT mutant with purple dots, the sextuple mutant with individual binding sites mutated with red dots, and the sextuple mutant with both binding sites mutated with gray dots. One-way ANOVA followed by Tukey’s multiple comparisons test; *P < 0.05, **P < 0.01, ***P < 0.001. n ≥ 9 for all genotypes.

WT, wild-type; a.u., arbitrary units. Scale bars 5 μm.