KIRREL3 localizes to the Golgi complex.

<p>Rat PNCs overexpressing the GFP-tagged KIRREL3 were immunostained with GFP antibody (green signal) (A) and the Golgi marker GS28 antibody (red signal, solid arrow) (B). Nuclei were stained with DAPI (blue signal, arrow head) (C). A distinct localized yellow signal (thin arrow) in the merged image (D) suggested the colocalization of KIRREL3 with the Golgi apparatus. Enlarged overlay images and individual red and green channels for a region of interest (ROI) are shown (E). The degree of overlap between the green and red signals was statistically analyzed (E) and expressed with Pearson’s correlation coefficient (PC) and Mander’s colocalization coefficients (M1 and M2). Bar, 20μm.</p

Nonmagnetic Quantum Emitters in Boron Nitride with Ultranarrow and Sideband-Free Emission Spectra

Hexagonal boron nitride (hBN) is an emerging material in nanophotonics and an attractive host for color centers for quantum photonic devices. Here, we show that optical emission from individual quantum emitters in hBN is spatially correlated with structural defects and can display ultranarrow zero-phonon line width down to 45 μeV if spectral diffusion is effectively eliminated by proper surface passivation. We demonstrate that undesired emission into phonon sidebands is largely absent for this type of emitter. In addition, magneto-optical characterization reveals cycling optical transitions with an upper bound for the g-factor of 0.2 ± 0.2. Spin-polarized density functional theory calculations predict possible commensurate transitions between like-spin electron states, which are in excellent agreement with the experimental nonmagnetic defect center emission. Our results constitute a step toward the realization of narrowband quantum light sources and the development of spin–photon interfaces within 2D materials for future chip-scale quantum networks

Western blot analysis of the Co-IP of KIRREL3-V5 with MAP1BLC1-FLAG (A1) and endogenous MAP1BLC1 (A2); KIRREL3-V5 with GFP-MYO16 (B), ATP1B1-FLAG (C), UFC1-FLAG (D), and GFP-KIRREL3 with SHMT2-FLAG (E).

<p>Lysates from HEK293H cells overexpressing the indicated expression constructs were incubated with anti-FLAG antibody (A1), anti-V5 antibody (A2, C, and D), and anti-GFP antibody (B, E), and precipitated with magnetic beads. Lysates from N2a cells overexpressing KIRREL3-V5 expression construct was incubated with anti-V5 antibody, and precipitated with magnetic beads (A2). Immunoprecipitates (lane IP, Co-IP) were analyzed by western blotting as indicated with anti-V5, anti-MAP1BLC1, anti-FLAG, and anti-GFP antibodies. Expression of all proteins was also analyzed in total lysates (lane L). MAP1BLC1-FLAG (green arrow) immobilizes KIRREL3-V5 (A1, lane IP, red arrow), but not LacZ-V5 (black arrow). KIRREL3-V5 (green arrow) and KIRREL3-ECD-V5 (brown arrow), but not LacZ-V5 (black arrow) immobilize endogenous MAP1BLC1 (A2, lane IP, red arrow). GFP-MYO16 (green arrow) immobilizes KIRREL3-V5 (B, lane IP, red arrow), KIRREL3-ECD-V5 (B, lane IP, brown arrow) but not LacZ-V5 (B, lane IP, black arrow). (C) KIRREL3-V5 (green arrow) but not LacZ-V5 (black arrow) immobilizes ATP1B1-FLAG (red arrow). (D) KIRREL3-V5 (green arrow) but not LacZ-V5 (black arrow) immobilizes UFC1-FLAG (red arrow). (E) GFP-KIRREL3 (green arrow) immobilizes SHMT2-FLAG (red arrow) but not BAP-FLAG (black arrow).</p

Individual yeast clones identified as potential interacting partners of KIRREL3-ECD and KIRREL3-ICD.

<p>(A) Positive control: yeast mating of [pGBKT7-p53] in AH109 and [pGADT7-T] in Y187. (B) Negative control: yeast mating of [pGBKT7- KIRREL3- ECD] in AH109 and [pGADT7-T] in Y187. (C) Yeast mating of [pGBKT7-KIRREL3-ECD] in AH109 and [pGADT7-MAP1BLC1] in Y187. (D) Yeast mating of [pGBKT7-KIRREL3 ECD] in AH109 and [pGADT7-MYO16] in Y187. (E) Negative control: yeast mating of [pGBKT7- KIRREL3-ICD] in AH109 and [pGADT7-T] in Y187. (F) Yeast mating of [pGBKT7-KIRREL3-ICD] in AH109 and [pGADT7-ATP1B1] in Y187. (G) Yeast mating of [pGBKT7-KIRREL3-ICD] in AH109 and [pGADT7-UFC1] in Y187. (H) Yeast mating of [pGBKT7-KIRREL3-ICD] in AH109 and [pGADT7-SHMT2] in Y187. Yeast mating was performed and cells were grown on—AHLT X-α-gal plates. Only clones with interacting proteins grow on—AHLT X-α-gal media and turn blue.</p

Schematic representation of the KIRREL3 domains.

<p>Five immunoglobulin domains (IgD), a signal peptide (SP) region, a transmembrane domain (TMD), and a PDZ- domain binding motif (PDZ-BD) are shown. ECD, extracellular domain; ICD, intracellular domain. The blue arrow indicates a potential cleavage site.</p

Autism and Intellectual Disability-Associated KIRREL3 Interacts with Neuronal Proteins MAP1B and MYO16 with Potential Roles in Neurodevelopment - Fig 4

<p>(A) KIRREL3-V5 (red, i) and MAP1BLC1-FLAG (green, ii) co-overexpressed in rat PNCs. (B) KIRREL3-V5 (red, i) and GFP-MYO16 (green, ii) co-overexpressed in rat PNCs. The overlapping signals of the two proteins appear as yellow/orange (Aiii, Biii) within the region of cytoplasm and in neurite processes (arrows). Enlarged overlay images and individual red and green channels for each ROIs (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0123106#sec009" target="_blank">Materials and Methods</a>) are shown (A-B, iv and v). The degree of colocalization between the red and green signals was statistically analyzed and expressed with Pearson’s correlation coefficient (PC) and Mander’s colocalization coefficients M1 and M2 (A-B, iv and v). M1 represents the fraction of KIRREL3-V5 (red signal) overlapped with MAP1BLC1-FLAG or GFP-MYO16 (green signal). M2 represents the fraction of MAP1BLC1-FLAG or GFP-MYO16 (green signal) overlapped with the KIRREL3-V5 (red signal). All calculations for Pearson’s and Mander’s coefficients were performed by the ImageJ version 1.45s visualization software with JACoP plugin. Bar, 20μm.</p

Image3_Classification and biomarker gene selection of pyroptosis-related gene expression in psoriasis using a random forest algorithm.TIFF

Background: Psoriasis is a chronic and immune-mediated skin disorder that currently has no cure. Pyroptosis has been proved to be involved in the pathogenesis and progression of psoriasis. However, the role pyroptosis plays in psoriasis remains elusive.Methods: RNA-sequencing data of psoriasis patients were obtained from the Gene Expression Omnibus (GEO) database, and differentially expressed pyroptosis-related genes (PRGs) between psoriasis patients and normal individuals were obtained. A principal component analysis (PCA) was conducted to determine whether PRGs could be used to distinguish the samples. PRG and immune cell correlation was also investigated. Subsequently, a novel diagnostic model comprising PRGs for psoriasis was constructed using a random forest algorithm (ntree = 400). A receiver operating characteristic (ROC) analysis was used to evaluate the classification performance through both internal and external validation. Consensus clustering analysis was used to investigate whether there was a difference in biological functions within PRG-based subtypes. Finally, the expression of the kernel PRGs were validated in vivo by qRT-PCR.Results: We identified a total of 39 PRGs, which could distinguish psoriasis samples from normal samples. The process of T cell CD4 memory activated and mast cells resting were correlated with PRGs. Ten PRGs, IL-1β, AIM2, CASP5, DHX9, CASP4, CYCS, CASP1, GZMB, CHMP2B, and CASP8, were subsequently screened using a random forest diagnostic model. ROC analysis revealed that our model has good diagnostic performance in both internal validation (area under the curve [AUC] = 0.930 [95% CI 0.877–0.984]) and external validation (mean AUC = 0.852). PRG subtypes indicated differences in metabolic processes and the MAPK signaling pathway. Finally, the qRT-PCR results demonstrated the apparent dysregulation of PRGs in psoriasis, especially AIM2 and GZMB.Conclusion: Pyroptosis may play a crucial role in psoriasis and could provide new insights into the diagnosis and underlying mechanisms of psoriasis.</p

Broadband Light Collection Efficiency Enhancement of Carbon Nanotube Excitons Coupled to Metallo-Dielectric Antenna Arrays

The realization of on-chip quantum networks ideally requires lossless interfaces between photons and solid-state quantum emitters. We propose and demonstrate on-chip arrays of metallo-dielectric antennas (MDA) that are tailored toward efficient and broadband light collection from individual embedded carbon nanotube quantum emitters by trapping air gaps on chip that form cavity modes. Scalable implementation is realized by employing polymer layer dry-transfer techniques that avoid solvent incompatibility issues, as well as a planar design that avoids solid-immersion lenses. Cryogenic measurements demonstrate 7-fold enhanced exciton intensity when compared to emitters located on bare wafers, corresponding to a light collection efficiency (LCE) up to 92% in the best case (average LCE of 69%) into a narrow output cone of ±15° that enables a priori fiber-to-chip butt coupling. The demonstrated MDA arrays are directly compatible with other quantum systems, particularly 2D materials, toward enabling efficient on-chip quantum light sources or spin-photon interfaces requiring unity light collection, both at cryogenic or room temperature

Workflow of our GC-MS based untargeted metabolomic and targeted analyses for biomarker discovery.

<p>The number of candidate metabolites analyzed at specific steps is shown in parenthesis.</p

Example of a retrieved EIC for valine.

<p>The inset in the top left shows the expected ratios for the fragments based on the library to guide the visual inspection. The doted vertical lines show the expected and estimated elution time of the analyte. Although, the background signal of 73 from other compounds is reflected in the apex score, its impact on the AUC is diminished by baseline correction.</p