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All isolated flukes were collected by pipette and preserved in saline (90°C) before being pooled in 1 flask and counted.
Thus, a total of 5 separate culture flasks were used for the biomass collection (1 flask each for days 0, 3, and 5; 1 flask for days 8, and 14; one flask for days 18 and 22), and each of these 5 flasks originated from a single 4-week-old B. braunii culture.
shRNA sequences: DBE: GCTCACCGCCATTCATGAAGG NDE: AACGTCACGGACAAGGCCAGA Nonsilencing: TCTCGCTTGGGCGAGAGTAAG Ash1L: GCTGGTCATTTATTGCTCAAT Suz12: TTCTACAAACAGCATACAG Lentiviruses Production and Transduction HEK293T cells were seeded in DMEM-HIGH supplemented with 10% FBS without antibiotics in T25 tissue culture flasks (1 flask per infection).
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SH-SY5Y cells were seeded at 2 × 106 cells/T25 flask.
Briefly, HeLa (ATCC CCL2) or NIH3T3 (ATCC CRL1658) cells were seeded at a density of 5×105/T75 flask or 2.5×105 cells/T25 flask.
The cells were washed and seeded in a 75 cm2 flask in fresh culture medium (3 4 Pups/per flask).
H292 cells were seeded into T75 flask and incubated for 18 h.
Briefly, the shM2-MDCK cells in a T25 cm2 flask were infected with H5N1 virus.
HeLa cells (4×106/T75 flask) were transfected with 10 µg of the ORF2-expressing constructs.
For each Huh7 cell line, 1.5×105 cells were seeded in one T75 flask.
For growth curves, 200,000 cells were replated in a 25-cm2 flask after each subculturing passage.
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