The sequence tag allowed us to deconvolute the sequences and, therefore, to relate the sequence to a specific mouse sample after Illumina sequencing.
Southern hybridization and PCR-based analyses of DNA from tissues of the resulting mice (sampled ∼1 month after TM treatment) confirmed the TM-induced, Cre-mediated excision of the Ate1flox allele in Ate1flox/− CaggCreER mice.
Hence, we monitored the disease-free survival of wt, TACI-Ig, Vav-Bcl2 and double-transgenic mice and took serum samples after 2, 8 or 12 months.
After washing with PBS, diluted mouse serum samples collected after each immunization were incubated at 37 °C for 1 h, followed by washing three times, 100 μL of horseradish peroxidase (HRP -conjugated anti-mouse antibody at different dilutions (1:100, 1:1000, and 1:10,000) were added to wells.
The titers of immunoglobulin G antibody specific to H7N9-M2e and H5N1-M2e were gradually increased in mouse serum samples collected after each vaccination, reaching very high levels (>1 10) 10 days after the last booster vaccination.
Furthermore, DNA extracted from splenic tissue samples after euthanizing the mice at sixteen weeks post-challenge also did not show any evidence of infection by PCR analysis (data not shown).
In studies using transgenic mice, Ad5 (10/0.1 mL per mouse) was injected intravenously into wild-type (WT) C57BL/6, GATA1 CAR, or GATA1 CR1 mice and blood sampled after 10, 30, 360, and 1440 minutes.
Mouse liver samples were obtained after killing and fixed in 4% paraformaldehyde, and then embedded in paraffin.
To identify candidate metabolites, mouse plasma samples were collected after continuous oral and subcutaneous exposure to DE-71, a widely used commercial pentabromodiphenyl ether product, for 34 days.
Preimmune serum samples or serum samples after the third immunization of individual mice were pooled for each group.
