Figure 3. FACT Subunits HMG-4 and SPT-16 Prevent Neuron Fate Induction in the Intestine(A) RNAi against
hmg-4 and
spt-16 allows
gcy-5::GFP induction in the intestine. Dashed lines indicate the outline of animals. Scale bars represent 20 um.(B) Quantification of animals showing
gcy-5::GFP in the intestine in
che-1oe + hmg-4RNAi or spt-16RNAi. n indicates the number of animals counted. Error barsrepresent SEM.(C) Detection of transcripts derived from neuronal genes in hmg-4RNAi and spt-16RNAi intestinal cells by smFISH. Individual mRNA molecules are visualized as reddots. Controls were treated with mock hybridizations. Dashed boxes indicate the magnified area. Scale bars represent 2 um.(D and E) Quantification of smFISH hybridization signals (red dots) per GFP-positive cells. Quantification of neuronal transcripts in the intestine upon (D) hmg-4RNAi and (E)
spt-16 RNAi. For each condition, 20 GFP-positive cells were counted. Ordinary one-way ANOVA was used for statistical analysis. **, p < 0.01,***, p < 0.001, ****, p < 0.0001. Error bars represent SD.(F and G) Immunostaining of CRISPR-tagged (F)
hmg-3 with HA and (G)
hmg-4 with HA. Dashed lines indicate the outline of the animals. Scale bars represent 20 um.(H) Anti-SPT-16 immunostaining shows expression in somatic tissues and germline. Dashed lines indicate the outline of the animals. Scale bars represent 20 um.(I) Tissue-specific FACT isoforms prevent induction of neuronal genes in the germline and intestine.See also Figure S2.