acdh-1 nascent RNAs are co-labeled by MCP::GFP and smFISH probes. (A) Representative images showing the production of nascent mRNAs (green) and protein of acdh-1 (magenta) at embryonic stages. BFP::LMN-1 labels the nuclear membrane. One embryo without DualTag knock-in for acdh-1 was shown as a negative control. Arrowheads: nascent RNA. The boxed region is magnified (2×). Scale bars: 10 μm. (B) Quantification of the number of active alleles of acdh-1 in A. For each group, 10 embryos were analyzed. The number of cells analyzed was indicated above each bar. (C) Representative images to show mRNA imaging by MCP::GFP and single-molecule fluorescence in situ hybridization (smFISH) in a control strain (the endogenous acdh-1 was not tagged with the DualTag). FISH probes specifically recognize the sequence of 24xMS2 (MS2 probe). Nuclei were stained by DAPI (blue). The boxed region is magnified (2×). Scale bars: 5 μm. (D) Quantification of MCP::GFP-positive spots in the control strain, in which the endogenous acdh-1 was not tagged with the DualTag. The negative control (NC) RNA FISH probe hybridizes with the transcripts of the bacterial DapB gene. n represents the number of embryos. (E) Representative images to show the co-labeling of MCP::GFP-positive acdh-1 RNA foci by smFISH using probes against DapB (NC) or MS2 sequence. Arrowheads: mRNA spots. The boxed region is magnified (2×) for NC group and (1.6×) for MS2 group. Scale bars: 5 μm. (F) Quantification of co-labeling of mRNAs by MCP::GFP and RNA FISH probes in E. n represents the number of embryos. Results are displayed as mean ± SEM. ****P < 0.0001 (two-tailed unpaired Student’s t test). (G) Confocal images to show the transcriptional levels of acdh-1 at various larval stages in the intestinal cells. Arrowheads: nascent RNA labeled by MCP::GFP. For L1 and L2 groups, the boxed regions are magnified (3.5×). For L3 and L4 groups, the boxed regions are magnified (3.3×). Scale bars: 5 μm. (H) Quantification of the number of active alleles of acdh-1 in G. The number of cells analyzed was indicated above each bar.
Figure 3.

acdh-1 nascent RNAs are co-labeled by MCP::GFP and smFISH probes. (A) Representative images showing the production of nascent mRNAs (green) and protein of acdh-1 (magenta) at embryonic stages. BFP::LMN-1 labels the nuclear membrane. One embryo without DualTag knock-in for acdh-1 was shown as a negative control. Arrowheads: nascent RNA. The boxed region is magnified (2×). Scale bars: 10 μm. (B) Quantification of the number of active alleles of acdh-1 in A. For each group, 10 embryos were analyzed. The number of cells analyzed was indicated above each bar. (C) Representative images to show mRNA imaging by MCP::GFP and single-molecule fluorescence in situ hybridization (smFISH) in a control strain (the endogenous acdh-1 was not tagged with the DualTag). FISH probes specifically recognize the sequence of 24xMS2 (MS2 probe). Nuclei were stained by DAPI (blue). The boxed region is magnified (2×). Scale bars: 5 μm. (D) Quantification of MCP::GFP-positive spots in the control strain, in which the endogenous acdh-1 was not tagged with the DualTag. The negative control (NC) RNA FISH probe hybridizes with the transcripts of the bacterial DapB gene. n represents the number of embryos. (E) Representative images to show the co-labeling of MCP::GFP-positive acdh-1 RNA foci by smFISH using probes against DapB (NC) or MS2 sequence. Arrowheads: mRNA spots. The boxed region is magnified (2×) for NC group and (1.6×) for MS2 group. Scale bars: 5 μm. (F) Quantification of co-labeling of mRNAs by MCP::GFP and RNA FISH probes in E. n represents the number of embryos. Results are displayed as mean ± SEM. ****P < 0.0001 (two-tailed unpaired Student’s t test). (G) Confocal images to show the transcriptional levels of acdh-1 at various larval stages in the intestinal cells. Arrowheads: nascent RNA labeled by MCP::GFP. For L1 and L2 groups, the boxed regions are magnified (3.5×). For L3 and L4 groups, the boxed regions are magnified (3.3×). Scale bars: 5 μm. (H) Quantification of the number of active alleles of acdh-1 in G. The number of cells analyzed was indicated above each bar.

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