Heatmap showing number of samples in which each of the total 976 proteins was identified in A549 exosomes.

Proteins common to all nine replicates are shown in blue (223). Proteins present in >1 replicate and <9 replicates are shown in grey (359) and proteins present only in 1 replicate are shown in red (394).</p

Variability in protein cargo detection in technical and biological replicates of exosome-enriched extracellular vesicles - Fig 2

Venn diagrams and tables showing the average number of proteins (in total and in percentage of the total) identified across three TRs and three BRs out of the total proteins detected in the three cell lines H1993 (886), A549 (976) and H1975 (879), respectively for (A) TRs (B) BRs. The proteins unique to the three replicates is shown in pink, the proteins shared by 2 replicates is shown in yellow, and the proteins common to three replicates is shown in blue.</p

Table shows one-way repeated measures ANOVA F-values and corresponding <i>p</i>-values for the levels of proteins identified in EE-EVs in 3 TRs and 3 BRs for H1993, A549 and H1975 cell lines.

Table shows one-way repeated measures ANOVA F-values and corresponding p-values for the levels of proteins identified in EE-EVs in 3 TRs and 3 BRs for H1993, A549 and H1975 cell lines.</p

Heatmap showing number of samples in which each of the total 879 proteins was identified in H1975 exosomes.

Proteins common to all nine replicates are shown in blue (108). Proteins present in >1 replicate and <9 replicates are shown in grey (305) and proteins present only in 1 replicate are shown in red (466).</p

Schematic representation of exosomal samples collection.

Exosomes were collected from three different cell lines at three different passages (A, B, C) to provide three biological replicates (BRs). The exosome lysate from each of them was further divided into three technical replicates (TRs). The nine samples were grouped so that the TRs were TR1: R1, R2, R3, TR2: R4, R5, R6, TR3: R7, R8, and R9. The BRs were BR1: R1, R4, R7, B2: R2, R5, R8, BR3: R3, R6, and R9.</p

Heatmap representing exosomal proteins identified in H1975 in order of decreasing levels in TRs (R1-R2-R3, R4-R5-R6, R7-R8-R9) and in BRs (R1-R4-R7, R2-R5-R6, R3-R6-R9).

Boxes highlighted in dark red represent the most abundant, while boxes in dark blue represent the least abundant proteins.</p

Heatmap representing exosomal proteins identified in H1993 in order of decreasing levels in TRs (R1-R2-R3, R4-R5-R6, R7-R8-R9) and in BRs (R1-R4-R7, R2-R5-R6, R3-R6-R9).

Boxes highlighted in dark red represent the most abundant, while boxes in dark blue represent the least abundant proteins.</p

Variance analysis showing quantitative variability in TRs and BRs of three H1993, A549, and H1975 cell line exosomes.

Variance analysis showing quantitative variability in TRs and BRs of three H1993, A549, and H1975 cell line exosomes.</p

Heatmap representing exosomal proteins identified in A549 in order of decreasing levels in TRs (R1-R2-R3, R4-R5-R6, R7-R8-R9) and in BRs (R1-R4-R7, R2-R5-R6, R3-R6-R9).

Boxes highlighted in dark red represent the most abundant, while boxes in dark blue represent the least abundant proteins.</p

Variance analysis showing qualitative variability in TRs and BRs of three H1993, A549, and H1975 cell line exosomes.

Variance analysis showing qualitative variability in TRs and BRs of three H1993, A549, and H1975 cell line exosomes.</p