Embryonic stem cell (ESC) culture conditions are important for maintaining long-term self-renewal, and they influence cellular pluripotency state. Here, we report single cell RNA-sequencing of mESCs cultured in three different conditions: serum, 2i, and the alternative ground state a2i. We find that the cellular transcriptomes of cells grown in these conditions are distinct, with 2i being the most similar to blastocyst cells and including a subpopulation resembling the two-cell embryo state. Overall levels of intercellular gene expression heterogeneity are comparable across the three conditions. However, this masks variable expression of pluripotency genes in serum cells and homogeneous expression in 2i and a2i cells. Additionally, genes related to the cell cycle are more variably expressed in the 2i and a2i conditions. Mining of our dataset for correlations in gene expression allowed us to identify additional components of the pluripotency network, including Ptma and Zfp640, illustrating its value as a resource for future discovery.

T helper type 2 (Th2) cells are important regulators of mammalian adaptive immunity and have relevance for infection, autoimmunity, and tumor immunology. Using a newly developed, genome-wide retroviral CRISPR knockout (KO) library, combined with RNA-seq, ATAC-seq, and ChIP-seq, we have dissected the regulatory circuitry governing activation and differentiation of these cells. Our experiments distinguish cell activation versus differentiation in a quantitative framework. We demonstrate that these two processes are tightly coupled and are jointly controlled by many transcription factors, metabolic genes, and cytokine/receptor pairs. There are only a small number of genes regulating differentiation without any role in activation. By combining biochemical and genetic data, we provide an atlas for Th2 differentiation, validating known regulators and identifying factors, such as Pparg and Bhlhe40, as part of the core regulatory network governing Th2 helper cell fates.

We apply analytic bootstrap techniques to the four-point correlator of fundamental fields in the Wilson-Fisher model. In an ϵ-expansion crossing symmetry fixes the double discontinuity of the correlator in terms of CFT data at lower orders. Large spin perturbation theory, or equivalently the recently proposed Froissart-Gribov inversion integral, then allows one to reconstruct the CFT data of intermediate operators of any spin. We use this method to compute the anomalous dimensions and OPE coefficients of leading twist operators. To cubic order in ϵ the double discontinuity arises solely from the identity operator and the scalar bilinear operator, making the computation straightforward. At higher orders the double discontinuity receives contributions from infinite towers of higher spin operators. At fourth order, the structure of perturbation theory leads to a proposal in terms of functions of certain degree of transcendentality, which can then be fixed by symmetries. This leads to the full determination of the CFT data for leading twist operators to fourth order.

We bound EFT coefficients appearing in 2 → 2 photon scattering amplitudes in four dimensions. After reviewing unitarity and positivity conditions in this context, we use dispersion relations and crossing symmetry to compute sum rules and null constraints. This allows us to derive new rigorous bounds on operators with four, six, and eight derivatives, including two-sided bounds on their ratios. Comparing with a number of partial UV completions, we find that some of our bounds are saturated by the amplitudes that arise from integrating out a massive scalar or axion, while others suggest the existence of unknown amplitudes.

A number of studies showed that the development and the lifespan of Caenorhabditis elegans is dependent on mitochondrial function. In this study, we addressed the role of mitochondrial DNA levels and mtDNA maintenance in development of C. elegans by analyzing deletion mutants for mitochondrial polymerase gamma (polg-1(ok1548)). Surprisingly, even though previous studies in other model organisms showed necessity of polymerase gamma for embryonic development, homozygous polg-1(ok1548) mutants had normal development and reached adulthood without any morphological defects. However, polg-1 deficient animals have a seriously compromised gonadal function as a result of severe mitochondrial depletion, leading to sterility and shortened lifespan. Our results indicate that the gonad is the primary site of mtDNA replication, whilst the mtDNA of adult somatic tissues mainly stems from the developing embryo. Furthermore, we show that the mtDNA copy number shows great plasticity as it can be almost tripled as a response to the environmental stimuli. Finally, we show that the mtDNA copy number is an essential limiting factor for the worm development and therefore, a number of mechanisms set to maintain mtDNA levels exist, ensuring a normal development of C. elegans even in the absence of the mitochondrial replicase.