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Additional file 2: Figure S2. A SH-SY5Y cell growth following BPTF KO shown by colony formation assay [mean, 114.330 vs. 81.330 vs. 45.000]. Error bars represent SEM. B SH-SY5Y cell migration following BPTF KO shown by RTCA. Error bars represent SD. The data are representative of at least three independent experiments. * P

Additional file 1: Figure S1. A GFP fluorescence imaging of 2 unique shRNA constructs targeting BPTF in SK-N-BE (2) cells 96 hours after lentiviral infection, when observed best transfection efficiency. Scale bar is 50 μm. B Quantitative of BPTF after 96h infection with BPTF knockdown lentiviral, as detected by RT-PCR [mean, 1.222 vs. 0.845 vs. 0.972]. C Sequencing results of the BPTF KO cell in SK-N-BE (2) compared with Ctrl. D Western blot of BPTF in SK-N-BE (2) cell with BPTF KO [mean, 0.386 vs. 0.000]. E, G Sequencing results of the BPTF KO cell in SH-SY5Y compared with Ctrl. F, H Representative western blot of BPTF in SH-SY5Y cell [mean, 0.442 vs. 0.000, D; 0.225 vs. 0.000, F] with BPTF KO. Error bars represent SEM. The data are representative of at least three independent experiments. * P

Additional file 3: Figure S3. A The STRING database hinted that TFAP4 was one of the higher-ranked related molecules associated with BPTF. B The GEO NB cohorts showed that the expression of BPTF and TFAP4 were significantly positively correlated (n=498). r=0.211, P

Bovine herpesvirus type 1 (BHV-1) is a neurotropic herpesvirus that causes infectious rhinotracheitis and vulvovaginitis in cattle. The virion host shutoff protein encoded by the BHV-1 UL41 gene is highly conserved in the Alphaherpesvirinae subfamily. This protein can degrade viral and host messenger RNA (mRNA) to interrupt host defense and facilitate the rapid proliferation of BHV-1. However, studies on the BHV-1 UL41 gene are limited, and BHV-1 defective virus construction using the CRISPR/Cas9 system is somewhat challenging. In this study, we rapidly constructed a BHV-1 UL41-deficient strain using the CRISPR/Cas9 system in BL primary bovine-derived cells. BHV-1 UL41-defective mutants were screened by Western blot analysis using specific polyclonal antibodies as the primary antibodies. During the isolation and purification of the defective strain, a mixed virus pool edited by an efficient single-guide RNA (sgRNA) showed a plaque number reduction. Viral growth property assessment showed that BHV-1 UL41 was dispensable for replication, but the UL41-defective strain exhibited early and slowed viral replication. Furthermore, the BHV-1 UL41-deficient strain exhibited enhanced sensitivity to temperature and acidic environments. The BHV-1 UL41-deficient strain regulated viral and host mRNA levels to affect viral replication.

Additional file 1: Figure S1. Pathological structure of each subtype of LM. Figure S2. Schematic diagram of the bioinformatics pipeline to analyze the targeted deep sequencing data. Figure S3. Transfection rate quantified by qPCR. Table S1: The coverage of each sample and average depth.

Objectives To investigate the genetic causes of hearing loss with enlarged vestibular aqueduct (EVA) in two children from unrelated two Chinese families. Methods Sanger sequencing of all coding exons in SLC26A4 (encoding Pendrin protein) was performed on the two patients, their sibling and parents respectively. To predict and visualize the potential functional outcome of the novel variant, model building, structure analysis, and in silico analysis were further conducted. Results The results showed that the proband from family I harbored a compound heterozygote of SLC26A4 c.1174A>T (p.N392Y) mutation and c.1181delTCT (p.F394del) variant in exon 10, potentially altering Pendrin protein structure. In family II, the proband was identified in compound heterozygosity with a known mutation of c.919-2A>G in the splice site of intron 7 and a novel mutation of c.1023insC in exon 9, which results in a frameshift and translational termination, consequently leading to truncated Pendrin protein. Sequence homology analysis indicated that all the mutations localized at high conservation sites, which emphasized the significance of these mutations on Pendrin spatial organization and function. Conclusion In summary, this study revealed two compound heterozygous mutations (c.1174A>T/c.1181delTCT; c.919- 2A>G/c.1023insC) in Pendrin protein, which might account for the deafness of the two probands clinically diagnosed with EVA. Thus this study contributes to improve understanding of the causes of hearing loss associated with EVA and develop a more scientific screening strategy for deafness.

High-density genetic linkage maps are particularly important for quantitative trait loci (QTL) mapping, genome assembly, and marker-assisted selection (MAS) in plants. In this study, a high-density genetic linkage map of sunflower (Helianthus annuus L.) was constructed using an F2 population generated from a cross between Helianthus annuus L. '86-1' and 'L-1-OL-1' via specific-locus amplified fragment sequencing (SLAF-seq). After sequence preprocessing, 530.50 M reads (105.60 Gb) were obtained that contained a total of 343,197 SLAFs, of which 39,589 were polymorphic. Of the polymorphic SLAFs, 6,136 were organized into a linkage map consisting of 17 linkage groups (LGs) spanning 2,221.86 cM, with an average genetic distance of 0.36 cM between SLAFs. Based on this high-density genetic map, QTL analysis was performed that focused on four sunflower phenotypic traits: oleic acid content (OAC), plant height (PH), head diameter (HD), and stem diameter (SD). Subsequently, for these four traits eight QTLs were detected that will likely be useful for increasing our understanding of genetic factors underlying these traits and for use in marker-assisted selection (MAS) for future sunflower breeding.