Amino acid differences between SD16 and SD16-MA.

<p>The amino acid location of mutations are numbered and indicated with arrowheads on the linear sequence. The locations of regions of protein binding, or functions are indicated with rectangles and are labeled with respect to interacting viral proteins. The PB1, NP and RNP ribonucleocapsid protein binding regions are in purple, green and yellow respectively; PB2 cap binding regions are in orange; HA receptor sites are in red.</p

DataSheet3_Zebrafish cobll1a regulates lipid homeostasis via the RA signaling pathway.ZIP

Background:The COBLL1 gene has been implicated in human central obesity, fasting insulin levels, type 2 diabetes, and blood lipid profiles. However, its molecular mechanisms remain largely unexplored.Methods:In this study, we established cobll1a mutant lines using the CRISPR/Cas9-mediated gene knockout technique. To further dissect the molecular underpinnings of cobll1a during early development, transcriptome sequencing and bioinformatics analysis was employed.Results:Our study showed that compared to the control, cobll1a−/− zebrafish embryos exhibited impaired development of digestive organs, including the liver, intestine, and pancreas, at 4 days post-fertilization (dpf). Transcriptome sequencing and bioinformatics analysis results showed that in cobll1a knockout group, the expression level of genes in the Retinoic Acid (RA) signaling pathway was affected, and the expression level of lipid metabolism-related genes (fasn, scd, elovl2, elovl6, dgat1a, srebf1 and srebf2) were significantly changed (p Conclusion:Our study suggest that the loss of cobll1a resulted in disrupted RA metabolism, reduced lipoprotein expression, and abnormal lipid transport, therefore contributing to lipid accumulation and deleterious effects on early liver development.</p

DataSheet1_Zebrafish cobll1a regulates lipid homeostasis via the RA signaling pathway.ZIP

Background:The COBLL1 gene has been implicated in human central obesity, fasting insulin levels, type 2 diabetes, and blood lipid profiles. However, its molecular mechanisms remain largely unexplored.Methods:In this study, we established cobll1a mutant lines using the CRISPR/Cas9-mediated gene knockout technique. To further dissect the molecular underpinnings of cobll1a during early development, transcriptome sequencing and bioinformatics analysis was employed.Results:Our study showed that compared to the control, cobll1a−/− zebrafish embryos exhibited impaired development of digestive organs, including the liver, intestine, and pancreas, at 4 days post-fertilization (dpf). Transcriptome sequencing and bioinformatics analysis results showed that in cobll1a knockout group, the expression level of genes in the Retinoic Acid (RA) signaling pathway was affected, and the expression level of lipid metabolism-related genes (fasn, scd, elovl2, elovl6, dgat1a, srebf1 and srebf2) were significantly changed (p Conclusion:Our study suggest that the loss of cobll1a resulted in disrupted RA metabolism, reduced lipoprotein expression, and abnormal lipid transport, therefore contributing to lipid accumulation and deleterious effects on early liver development.</p

Viral growth kinetics of SD16 and SD16-MA in MDCK and A549 cells.

<p>Confluent MDCK (A) or A549 (B) cells were infected with H9N2 viruses at an MOI of 0.01. Virus yields at 12, 24, 36, 48, and 60 hpi were titrated in MDCK cells. Each data point represents the mean virus yield from three individually infected wells ± SD.</p

Table1_Zebrafish cobll1a regulates lipid homeostasis via the RA signaling pathway.pdf

Background:The COBLL1 gene has been implicated in human central obesity, fasting insulin levels, type 2 diabetes, and blood lipid profiles. However, its molecular mechanisms remain largely unexplored.Methods:In this study, we established cobll1a mutant lines using the CRISPR/Cas9-mediated gene knockout technique. To further dissect the molecular underpinnings of cobll1a during early development, transcriptome sequencing and bioinformatics analysis was employed.Results:Our study showed that compared to the control, cobll1a−/− zebrafish embryos exhibited impaired development of digestive organs, including the liver, intestine, and pancreas, at 4 days post-fertilization (dpf). Transcriptome sequencing and bioinformatics analysis results showed that in cobll1a knockout group, the expression level of genes in the Retinoic Acid (RA) signaling pathway was affected, and the expression level of lipid metabolism-related genes (fasn, scd, elovl2, elovl6, dgat1a, srebf1 and srebf2) were significantly changed (p Conclusion:Our study suggest that the loss of cobll1a resulted in disrupted RA metabolism, reduced lipoprotein expression, and abnormal lipid transport, therefore contributing to lipid accumulation and deleterious effects on early liver development.</p

DataSheet2_Zebrafish cobll1a regulates lipid homeostasis via the RA signaling pathway.ZIP

Background:The COBLL1 gene has been implicated in human central obesity, fasting insulin levels, type 2 diabetes, and blood lipid profiles. However, its molecular mechanisms remain largely unexplored.Methods:In this study, we established cobll1a mutant lines using the CRISPR/Cas9-mediated gene knockout technique. To further dissect the molecular underpinnings of cobll1a during early development, transcriptome sequencing and bioinformatics analysis was employed.Results:Our study showed that compared to the control, cobll1a−/− zebrafish embryos exhibited impaired development of digestive organs, including the liver, intestine, and pancreas, at 4 days post-fertilization (dpf). Transcriptome sequencing and bioinformatics analysis results showed that in cobll1a knockout group, the expression level of genes in the Retinoic Acid (RA) signaling pathway was affected, and the expression level of lipid metabolism-related genes (fasn, scd, elovl2, elovl6, dgat1a, srebf1 and srebf2) were significantly changed (p Conclusion:Our study suggest that the loss of cobll1a resulted in disrupted RA metabolism, reduced lipoprotein expression, and abnormal lipid transport, therefore contributing to lipid accumulation and deleterious effects on early liver development.</p

Virulence and death pattern of mice infected with different H9N2 viruses.

<p>Five-week-old BALB/c mice (five/group) were inoculated intranasally with different H9N2 viruses, SD16 (A), SD16-MA (B), SD16:MA PB2 (C), or SD16-M147L/E627K (D). Doses of 10⁴ to 10⁶ pfu (A) or 10² to 10⁵ pfu (B, C, D) were used.</p

DataSheet5_Zebrafish cobll1a regulates lipid homeostasis via the RA signaling pathway.docx

Background:The COBLL1 gene has been implicated in human central obesity, fasting insulin levels, type 2 diabetes, and blood lipid profiles. However, its molecular mechanisms remain largely unexplored.Methods:In this study, we established cobll1a mutant lines using the CRISPR/Cas9-mediated gene knockout technique. To further dissect the molecular underpinnings of cobll1a during early development, transcriptome sequencing and bioinformatics analysis was employed.Results:Our study showed that compared to the control, cobll1a−/− zebrafish embryos exhibited impaired development of digestive organs, including the liver, intestine, and pancreas, at 4 days post-fertilization (dpf). Transcriptome sequencing and bioinformatics analysis results showed that in cobll1a knockout group, the expression level of genes in the Retinoic Acid (RA) signaling pathway was affected, and the expression level of lipid metabolism-related genes (fasn, scd, elovl2, elovl6, dgat1a, srebf1 and srebf2) were significantly changed (p Conclusion:Our study suggest that the loss of cobll1a resulted in disrupted RA metabolism, reduced lipoprotein expression, and abnormal lipid transport, therefore contributing to lipid accumulation and deleterious effects on early liver development.</p

Replication and pathogenicity of mutants in mice.

a<p>Five-week-old female BALB/c mice were inoculated intranasally with 10⁵ pfu of viruses. Three mice from each group were killed at 3dpi and virus titers in lung were determined.</p>b<p>number of mice infected/number of mice inoculated.</p

Cytokine responses in lungs of infected mice.

<p>Groups of three BALB/c mice were inoculated intranasally with 10⁵ pfu of SD16 or SD16-MA virus. The lungs of mice were collected at 3 dpi. The concentrations of various cytokines in lung homogenates were measured by a protein array analysis with Bio-Plex Mouse Cytokine 8-Plex. Each value represents mean cytokine concentration of three mice from each infected group ± SD. *, <i>P</i><0.05 compared with the value of SD16 infected mice.</p