- WBPaper00037901:GLD-1_mRNA_targets
Authors calculated the mean IP enrichment from the two analyses and given a cutoff of three-fold, they identified 948 reproducibly enriched mRNAs (14.2%) from of a total of 6635 detected by the array.
948 reproductively enriched mRNAs that co-immunoprecipitate with GLD-1. To identify GLD-1 mRNA targets, authors performed immunoprecipitation (IP) of GLD-1, followed by microarray analysis of the co-IPed mRNAs (RIP-chip). Extracts from young adult transgenic worms expressing a rescuing FLAG and GFP-tagged GLD-1, hereafter referred to as tagged GLD-1, were subjected to IP in triplicate with anti-FLAG (aFLAG IP) or anti-MYC (aMYC IP) antibodies as controls. Comparison of aFLAG IP versus aMYC IP to input revealed a large population of GLD-1-associated transcripts. Authors additionally performed complementary aFLAG IPs upon worms expressing either tagged GLD-1(GGF IP) or non-tagged GLD-1(N2 IP). Comparing transcript 'IP-enrichment values' from both approaches revealed a correlation of 0.96, which indicated high reproducibility of GLD-1 association with specific mRNAs even on a quantitative level.
- WBPaper00031252:AIN-1_IP_enriched
Signals from replicates of total worm lysates from wt and strains containing the ain-2::gfp or the ain-2 promoter::gfp transgene were mean normalized and averaged respectively to generate standard profiles of gene expression in these worm strains. Authors then calculated the ratio of signal of each gene from each IP sample to the standard gene expression profile of the corresponding worm strain. Based on this ratio, a percentile rank of each gene relative to all genes in each IP replicate was calculated. The percentile ranks in the three replicates of each IP were averaged. Student t test was utilized to determine if the average percentile ranks of enrichment of individual genes were significantly higher (p value) than the mean enrichment of all genes in the IP samples. To determine the AIN-1 or AIN-2 associated genes, we used the following criteria: (1) average percentile ranks of enrichment is greater than the mean enrichment of all genes in AIN-1 or AIN-2 IP with p < 0.01; (2) average signal in AIN-1 or AIN-2 IP replicates is greater than the background signal (including 2X standard deviation (SD)) (Background signal and SD were calculated based on signals from empty spots on each microarray); (3) criteria 1 is not be satisfied for the same gene in the corresponding control IP.
mRNAs that were significantly enriched in the AIN-1 immunoprecipitation samples, compared to the control total mRNAs in the input extracts (p < 0.01).
- WBPaper00031252:AIN-2_IP_enriched
Signals from replicates of total worm lysates from wt and strains containing the ain-2::gfp or the ain-2 promoter::gfp transgene were mean normalized and averaged respectively to generate standard profiles of gene expression in these worm strains. Authors then calculated the ratio of signal of each gene from each IP sample to the standard gene expression profile of the corresponding worm strain. Based on this ratio, a percentile rank of each gene relative to all genes in each IP replicate was calculated. The percentile ranks in the three replicates of each IP were averaged. Student t test was utilized to determine if the average percentile ranks of enrichment of individual genes were significantly higher (p value) than the mean enrichment of all genes in the IP samples. To determine the AIN-1 or AIN-2 associated genes, we used the following criteria: (1) average percentile ranks of enrichment is greater than the mean enrichment of all genes in AIN-1 or AIN-2 IP with p < 0.01; (2) average signal in AIN-1 or AIN-2 IP replicates is greater than the background signal (including 2X standard deviation (SD)) (Background signal and SD were calculated based on signals from empty spots on each microarray); (3) criteria 1 is not be satisfied for the same gene in the corresponding control IP.
mRNAs that were significantly enriched in the AIN-2 immunoprecipitation samples, compared to the control total mRNAs in the input extracts (p < 0.01).
- WBPaper00061479:HDA-1_interacting_protein
Proteins either absent in N2 or with more than twofold of the spectra counts in the hda-1::gfp IP compared to those in N2 are shown.
Proteins interacting with HDA-1
- WBPaper00062103:neuron_enriched
DESEQ2, fold change > 2 and FDR < 0.01.
Neuronally enriched transcripts according to a comparison of neuronal nuclei IP samples to total nuclei using isolation of nuclei from tagged specific cell types (INTACT) technology.