Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Validation of small extracellular vesicles. ( A ) The workflow of the isolation of small extracellular vesicles (sEVs) from the plasma of healthy controls (HCs) and recovered COVID-19 patients (RCs), including recovered asymptomatic patients (RAs), recovered moderate patients (RMs), and recovered severe and critical patients (RSs). ( B ) Representative transmission electron microscopy of the morphology of sEVs using commercial Exosome kit from RAs, RMs, RSs, and HCs. Scale bar: 200 nm. ( C ) Nanoparticle tracking analysis of sEVs from the plasma of RAs, RMs, RSs, and HCs. The detailed data were presented in Tables and . ( D ) The size distribution of the purified sEVs isolated from plasma of HCs, RAs, RMs, and RSs. ( E ) Western blotting indicates the expression levels of CD9, CD63, TSG101, Calnexin, GM130, GRP94, and Lamin B1 in cells, RAs, RMs, RSs, and HCs. Among them, cells were selected as control. The gels were cropped from different parts and automatically exposed using BIORAD. Cropped blots were displayed alongside their corresponding original blots in Fig. .

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques: Biomarker Discovery, Isolation, Clinical Proteomics, Transmission Assay, Electron Microscopy, Purification, Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: ( A ) Percentage of identified sEVs cargo lipid classes and subclasses. ( B ) Relative abundance (%) of sEVs lipids subclasses in recovered asymptomatic patients (RAs), recovered moderate patients (RMs), recovered severe/critical patients (RSs), and healthy controls (HCs). ( C – E ) The level of total lipids and lipids subclasses diacylglycerols (DAGs) and phosphatidylcholines (PCs) in RAs, RMs, RSs, and HCs. Data were displayed as boxplots with median and range and each dot represents an individual: HC (Pink), RA (Green), RM (Red), RS (Blue). p-value: *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques:

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Lipid profiles of sEVs isolated from HCs, RAs, RMs, and RSs plasma. ( A – C ) Orthogonal partial least square discriminant analysis score plot in recovered asymptomatic patients (RAs), recovered moderate patients (RMs), and recovered severe/critical patients (RSs) compared to HCs. ( D ) The heatmap shows significantly altered lipids in the sEVs isolated from the plasma of RAs, RMs, RSs, and HCs. Only differential lipids with p < 0.05 were displayed, and the color shades indicated the levels of lipids. Yellow and blue are indicative of relatively higher and lower levels, respectively.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques: Isolation, Clinical Proteomics

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: ( A – C ) Volcano plots show the sEVs differential lipids in RAs ( A ), RMs ( B ), or RSs ( C ) compared with HCs. The x-axis was the value of log2(FC), FC indicates the ratio of the mean level of the sEVs lipid in RAs, RMs, or RSs to the mean value of HCs, y-axis was − lg(p-value). Blue represented downregulation with p < 0.05, red represented upregulation, and grey denoted no change. ( D ) Venn diagram shows the number of significantly altered lipids in RAs, RMs, and RSs relative to HCs. ( E ) Statistical analysis of FFA(22:5) was displayed as bar graphs in sEVs of HCs, RAs, RMs, and RSs. The significance of comparisons was determined by the Kruskal–Wallis test. Data were displayed as boxplots with median and range and each dot represents an individual: HC (Pink), RA (Green), RM (Red), RS (Blue). Lipids quantitated using targeted lipidomics were presented in nanograms of lipids per milliliter (ng/mL) sEVs. p-value: *p < 0.05; **p < 0.01; ***p < 0.001. RAs recovered asymptomatic patients, RMs recovered moderate patients, RSs recovered severe/critical patients.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques:

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Distinct lipid profiles in RCs (RAs, RMs, and RSs) and KEGG enrichment analysis. ( A – C ) Volcano plots presenting differential lipids in RM vs. RA, RS vs. RA, and RS vs. RM. The x-axis is the log2(FC) value, and the y-axis is the − lg (p-value). Blue represented downregulation with p < 0.05, red represented upregulation, and grey denoted no change. ( D ) Venn plot showing the differential lipids among diverse RC groups (RAs, RMs, and RSs). The blue, red, and green circles represent RM vs. RA, RS vs. RA, and RS vs. RM groups. ( E ) Statistical analysis of DAG(16:1/18:0) is displayed as the box plot graph. The significance of comparisons was determined by the Kruskal–Wallis test. Lipids quantitated using targeted lipidomics were presented in nanomoles of lipids per milliliter (ng/mL) sEVs. p-value: *p < 0.05; **p < 0.01; ***p < 0.001. ( F ) Lipidomic KEGG enrichment analysis of differential lipids in RCs relative to HCs. The pathway enrichment analysis calculates the p-value and pathway impact, respectively. The node color is based on the p-value, and the node radius is determined based on pathway impact values.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques:

Journal: Scientific Reports

Article Title: Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

doi: 10.1038/s41598-023-43189-5

Figure Lengend Snippet: Correlation of plasma sEVs lipids with clinical indices. ( A – C ) Correlation plots illustrate Spearman correlations between clinical indices with differential lipids identified in RAs ( A ), RMs ( B ), and RSs ( C ) relative to HCs in 102 recruiters. Only correlation with p < 0.05 was indicated with colored squares and marked with stars (*). Positive and negative correlations were shown in blue and red, respectively, with sizes of squares representing the magnitude of the correlations. *p < 0.05; ** p < 0.01; ***p < 0.001. RAs recovered asymptomatic patients, RMs recovered moderate patients, RSs recovered severe/critical patients.

Article Snippet: Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies , .

Techniques: Clinical Proteomics

Journal: Ecotoxicology and environmental safety

Article Title: Butylparaben disordered intestinal homeostasis in Chinese striped-necked turtles (Mauremys sinensis).

doi: 10.1016/j.ecoenv.2023.115193

Figure Lengend Snippet: Fig. 6. The observation of neutrophil and NK cells through immunofluorescence assay. Neutrophils (MPO positive cells); NK cells (CD56 positive cells). BuP-I: 5 μg/L BuP group; BuP-II: 50 μg/L BuP group; BuP-III: 500 μg/L BuP group.

Article Snippet: Subsequently, the sections were incubated with the rabbit anti-MPO and anti-CD56 antibodies (1: 200 dilution) (BOSTER Biological Technology, Wuhan, China).

Techniques: Immunofluorescence

Journal: bioRxiv

Article Title: Upregulation of FasII underlies synergistic neuropathological and behavioral defects in a Drosophila model of myotonic dystrophy

doi: 10.1101/2024.05.26.595976

Figure Lengend Snippet: a, b Representative semi-quantitative RT-PCR of fasII in Drosophila larval (a) CNS and (b) BWM. c Quantification of a . n = 3 for both genotypes. d Quantification of b . n = 3 for both genotypes. e, f Representative slot blots of FasII in adult Drosophila (e) head and (f) muscle. g Quantification of e . n = 3 for both genotypes. h Quantification of f . n = 3 for both genotypes. i, j Representative slot blots of NCAM1 in mouse (i) head and (j) muscle. k Quantification of i . n = 3 for both genotypes. l Quantification of j . n = 3 for both genotypes. m Quantified slot blot data of NCAM1 in human frontal cortex. n = 5 for control. n = 10 for DM1 patients. n Quantitative dot blot data of NCAM1 in human transdifferentiated myotube. n = 4 for both control and DM1 patients. Histograms depicts mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Membranes were blocked in 5% Blotto (Santa Cruz Biotech, sc-2324) diluted in 1X TBS-T (10 mM Tris-HCl, 0.15 M NaCl, 0.05% Tween 20) for 1 hour at room temperature, and incubated overnight at 4°C with primary anti-NCAM1 antibody (GeneTex, GTX111684) diluted 1:10,000 in 5% Blotto.

Techniques: Quantitative RT-PCR, Dot Blot, Control, Quantitative Dot Blot