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The survival fraction of AL cells, assayed by PI-Flow Cytometry, manifested a PI-negative (indication of living cells) fraction of 12.7 ± 3.37% at 20% (v/v) glycerol pre-treatment.
The aqueous-octane culture system was maintained in operation for 24 h, then the volume of the aqueous phase corresponding to a final 0.05 OD was used as the inoculum for a second batch reactor (taking into account the viable cells fraction).
Similar to the isolation of PDCs for B cells, single-cell suspension was incubated for 20 min at 4°C with Biotin-Antibody Cocktail (CD43, CD4 and Ter119) and Anti-Biotin MicroBeads, and then the labeled cells were depleted with LS column in order to obtain the B cells fraction.
Hence, we break the pro-B and pre-B subsets into two subsets each: Bor for small resting pro-B cells (Hardy's fractions A through C) and Boc for large cycling pro-B cells (part of Hardy's fraction C); similarly, Bec for large cycling pre-B cells (the remainder of fraction C) and Ber for small resting pre-B cells (fraction D).
The purity of the NK cells fraction was ≥ 98%.
We compared the transcriptome of senescent yeast mother cells (fraction V) with young daughter cells (fraction II) after separating them by elutriation centrifugation [ 2].
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The table presents the VAFs in each cell fraction.
(g) Amplicon sequencing results on FACS-sorted cells show the VAFs in each cell fraction.
The live cell fraction was gated to analyze the early phase of apoptotic cell death.
(b) Cell fraction of cytosol (C) and nucleus (N) was analysed by western blot.
The mononuclear cell fraction was isolated and culture expanded.
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