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A similar analysis is applied in Figure 2C to a different neuron sample.
The unlabeled cell fraction for the POMC-GFP animals largely contains the mature granule cell population and was used as the "mature neuron" sample.
These significant effects were present in a high proportion of the single neuron sample despite the small number of randomly recorded neurons included in the sample at each site, an average of 2.5 neurons (range 1 4).
RNAseq data sets were downloaded from the RNAseq Atlas (http://medicalgenomics.org/rna_seq_atlas), and the following tissues were included: colon, liver, ovary and hypothalamus (matched with a neuron sample for methylation).
Gpr17, Mbp, Mobp, and Plp1 are all highly expressed in oligodendrocytes and are also found differentially expressed in the wildtype AGRP neuron sample from microarray data from Ren et al. (2012).
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In a neuronal population each neuron samples from a unique retinal position, which enables the encoding of temporal changes at every retinal position.
The differential expression for these primarily non-neuronal markers in our AGRP and POMC neuron samples relative to purified samples of these non-neuronal cell types was at least 300-fold (range: 318- to 8904-fold), indicating a high level of sample purity.
Expression of a list of well-acknowledged neuronal markers could clearly segregate blood samples from single neuron samples (Fig. 2E).
When we evaluated expression of 1479 genes included in Dim1 by Darmanis et al. in our single neuron samples, we found a subset of the genes had enriched expression in our "matured" neuronal samples (Fig. S2).
For each gene pair, comparison was made across three undifferentiated and three P19 neuron samples.
Chromatin was extracted from three P19 stem cell samples and three P19 neuron samples following the manufacturer's instructions (EZ-ChIP kit; Upstate Biotechnology, Charlottesville, VA).
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