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( A–I ) Analysis of ABs, MVs, and EXOs isolated from human EF samples: morphology by TEM ( A , D , and G ), size distribution by DLS ( B , E , and H ), and protein marker expression by Western blotting ( C , F , and I ). TEM images obtained using two different protocols for an external (deposition processing, upper images) or internal (ultrathin slide processing, lower images) view of EVs. Size distribution analyzed in a single EF sample by DLS during the receptive phase for ( B ) ABs, ( E ) MVs, and ( H ) EXOs. Graphs show the average size distribution and percentage of total particles contained within the populations. Specific protein markers analyzed by Western blotting for ( C ) ABs, ( F ) MVs, and ( I ) EXOs. Analyzed markers (and associated molecular mass) were calnexin (90–100 kDa), calreticulin (60 kDa), VDAC1 (31 kDa), <t>ARF6</t> (18 kDa), CD9 (24 kDa), CD63 (30–60 kDa), and TSG101 (45–50 kDa). ( J and K ) Particle concentration and size distribution measured by NTA for ( J ) MVs and ( K ) EXOs secreted throughout the menstrual cycle. One-way ANOVA and Kruskal-Wallis rank sum tests performed to compare the distinct menstrual cycle phases - no significant differences were observed. Figure 1—source data 1. Unedited blots for .
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( A–I ) Analysis of ABs, MVs, and EXOs isolated from human EF samples: morphology by TEM ( A , D , and G ), size distribution by DLS ( B , E , and H ), and protein marker expression by Western blotting ( C , F , and I ). TEM images obtained using two different protocols for an external (deposition processing, upper images) or internal (ultrathin slide processing, lower images) view of EVs. Size distribution analyzed in a single EF sample by DLS during the receptive phase for ( B ) ABs, ( E ) MVs, and ( H ) EXOs. Graphs show the average size distribution and percentage of total particles contained within the populations. Specific protein markers analyzed by Western blotting for ( C ) ABs, ( F ) MVs, and ( I ) EXOs. Analyzed markers (and associated molecular mass) were calnexin (90–100 kDa), calreticulin (60 kDa), VDAC1 (31 kDa), <t>ARF6</t> (18 kDa), CD9 (24 kDa), CD63 (30–60 kDa), and TSG101 (45–50 kDa). ( J and K ) Particle concentration and size distribution measured by NTA for ( J ) MVs and ( K ) EXOs secreted throughout the menstrual cycle. One-way ANOVA and Kruskal-Wallis rank sum tests performed to compare the distinct menstrual cycle phases - no significant differences were observed. Figure 1—source data 1. Unedited blots for .
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( A–I ) Analysis of ABs, MVs, and EXOs isolated from human EF samples: morphology by TEM ( A , D , and G ), size distribution by DLS ( B , E , and H ), and protein marker expression by Western blotting ( C , F , and I ). TEM images obtained using two different protocols for an external (deposition processing, upper images) or internal (ultrathin slide processing, lower images) view of EVs. Size distribution analyzed in a single EF sample by DLS during the receptive phase for ( B ) ABs, ( E ) MVs, and ( H ) EXOs. Graphs show the average size distribution and percentage of total particles contained within the populations. Specific protein markers analyzed by Western blotting for ( C ) ABs, ( F ) MVs, and ( I ) EXOs. Analyzed markers (and associated molecular mass) were calnexin (90–100 kDa), calreticulin (60 kDa), VDAC1 (31 kDa), <t>ARF6</t> (18 kDa), CD9 (24 kDa), CD63 (30–60 kDa), and TSG101 (45–50 kDa). ( J and K ) Particle concentration and size distribution measured by NTA for ( J ) MVs and ( K ) EXOs secreted throughout the menstrual cycle. One-way ANOVA and Kruskal-Wallis rank sum tests performed to compare the distinct menstrual cycle phases - no significant differences were observed. Figure 1—source data 1. Unedited blots for .
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( A–I ) Analysis of ABs, MVs, and EXOs isolated from human EF samples: morphology by TEM ( A , D , and G ), size distribution by DLS ( B , E , and H ), and protein marker expression by Western blotting ( C , F , and I ). TEM images obtained using two different protocols for an external (deposition processing, upper images) or internal (ultrathin slide processing, lower images) view of EVs. Size distribution analyzed in a single EF sample by DLS during the receptive phase for ( B ) ABs, ( E ) MVs, and ( H ) EXOs. Graphs show the average size distribution and percentage of total particles contained within the populations. Specific protein markers analyzed by Western blotting for ( C ) ABs, ( F ) MVs, and ( I ) EXOs. Analyzed markers (and associated molecular mass) were calnexin (90–100 kDa), calreticulin (60 kDa), VDAC1 (31 kDa), ARF6 (18 kDa), CD9 (24 kDa), CD63 (30–60 kDa), and TSG101 (45–50 kDa). ( J and K ) Particle concentration and size distribution measured by NTA for ( J ) MVs and ( K ) EXOs secreted throughout the menstrual cycle. One-way ANOVA and Kruskal-Wallis rank sum tests performed to compare the distinct menstrual cycle phases - no significant differences were observed. Figure 1—source data 1. Unedited blots for .

Journal: eLife

Article Title: Vertical transmission of maternal DNA through extracellular vesicles associates with altered embryo bioenergetics during the periconception period

doi: 10.7554/eLife.88008

Figure Lengend Snippet: ( A–I ) Analysis of ABs, MVs, and EXOs isolated from human EF samples: morphology by TEM ( A , D , and G ), size distribution by DLS ( B , E , and H ), and protein marker expression by Western blotting ( C , F , and I ). TEM images obtained using two different protocols for an external (deposition processing, upper images) or internal (ultrathin slide processing, lower images) view of EVs. Size distribution analyzed in a single EF sample by DLS during the receptive phase for ( B ) ABs, ( E ) MVs, and ( H ) EXOs. Graphs show the average size distribution and percentage of total particles contained within the populations. Specific protein markers analyzed by Western blotting for ( C ) ABs, ( F ) MVs, and ( I ) EXOs. Analyzed markers (and associated molecular mass) were calnexin (90–100 kDa), calreticulin (60 kDa), VDAC1 (31 kDa), ARF6 (18 kDa), CD9 (24 kDa), CD63 (30–60 kDa), and TSG101 (45–50 kDa). ( J and K ) Particle concentration and size distribution measured by NTA for ( J ) MVs and ( K ) EXOs secreted throughout the menstrual cycle. One-way ANOVA and Kruskal-Wallis rank sum tests performed to compare the distinct menstrual cycle phases - no significant differences were observed. Figure 1—source data 1. Unedited blots for .

Article Snippet: Membranes were blocked in 5% skim milk in 1% Tween-20 in PBS (PBS-T) and incubated overnight at 4 °C with the following primary antibodies: rabbit anti-calnexin (1:1000; ADI-SPA-865; Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-calreticulin (1:1000; ab22683; Abcam, Cambridge, UK), rabbit anti-VDAC1 (1:1000; ab154856; Abcam), rabbit anti-ARF6 (1:1000; ab77581; Abcam), mouse anti-CD63 (1:1000; ab59479; Abcam), rabbit anti-CD9 (1:1000; ab92726; Abcam), and rabbit anti-TSG101 (1:1000; 125011; Abcam).

Techniques: Isolation, Marker, Expressing, Western Blot, Concentration Assay