8/21/2023 0 Comments Per2 snapgene![]() **p ≤ 0.01, ***p ≤ 0.001, two-way analysis of variance (ANOVA) with Bonferroni’s correction for multiple comparisons. The average fold enrichment in QK159 samples was then normalized to the average fold enrichment in N2 samples within each trial. Values for the amplicon of interest in QK159 and N2 (wild type) were first normalized to the respective input. Values represent the mean ± standard error of the mean (SEM) of three independent trials, each of which included three technical replicates. The promoter of col-19, which had no detectable NHR-23 ChIP-seq peaks, was used as a negative control. ( B, C) ChIP-qPCR analysis of NHR-23::3xFLAG enrichment at the let-7 promoter in 元- and L4-stage wild-type (untagged control) and nhr-23::3xflag larvae. Bottom: The NHR-23 ChIP-seq peaks reported by the modENCODE Consortium are indicated. Dotted line represents the amplicon quantified by ChIP-qPCR. TSS indicates the major transcriptional start site for pri- let-7 ( Kai et al., 2013). Top: Mature let-7 (magenta) pri-let-7 (black) upstream DNA sequences (dark gray) present in the let-7p::gfp transcriptional reporter ( Kai et al., 2013) the minimal seam-specific enhancer (light gray) (MP) ( Johnson et al., 2003) consensus ROREs (brown). elegans (top) and corresponding NHR-23 ChIP-seq peaks (bottom). Supplementary file 1 includes the active, lethargic, and W–W intervals of these eight cohorts, as well as additional cohorts of both let-7 family mutants and nhr-23 knockdowns. ****p ≤ 0.0001, *p ≤ 0.05 ordinary one-way ANOVA with Bonferroni’s correction for multiple comparisons. ( C) Same as B, except that the actograms depict animals observed from emergence in 元 onward. ****p ≤ 0.0001 ordinary one-way analysis of variance (ANOVA) with Bonferroni’s correction for multiple comparisons. Relevant scoring rubrics are further defined in Results and Materials and methods. The wake-to-wake time intervals (W–W) are indicated. See key at the bottom of Figure 2 for details activity is designated as low (L), medium (M), or high (H) Mlt and Rup indicate molting and rupture phenotypes, respectively. In addition, the molting-defective phenotype is labeled only when first detected. ![]() The records within each column correspond to a single worm. ![]() Each chart combines records from two independent trials. ( B) Actograms depict the behavior, life stage, and phenotype of worms observed at 1h intervals from emergence in L4 onward. ( A) Stages of the life cycle (left) and the molting cycle (right) of C.
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