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In control group, all the mice were unimmunized, including their natal offspring.
In three of the six experimental groups, the female mice were unimmunized, but their offspring were immunized with 1.0 μg, 0.1 μg and 0.01 μg of inactivated influenza virus A/PR/8/34 vaccine at age of 1 week respectively, and boosted 3 weeks later with the vaccine at the same dose as primed.
In two of the four experimental groups, the female mice were unimmunized, but their offspring were immunized at age of 1 week with 30 μg of HA DNA and 30 μg of NA DNA, respectively, and boosted 3 weeks later with the same DNA vaccine at the same dose as primed.
However, when the female mice were unimmunized, the offspring immunized with HA DNA and NA DNA showed the survival rates of 71.4% and 100%, respectively (Table 2 and Figure 1B), and they had significantly lower lung virus titers than the offspring in control group, and lower titers than those born to immunized mothers as well.
However, when the female mice were unimmunized, their offspring immunized with 1.0 μg, 0.1 μg and 0.01 μg of inactivated influenza vaccine had the survival rates of 100%, 100%and83.3%3% respectively (Table 1 and Figure 1A), with significantly lower lung virus titers than offspring in control group.
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In control group, both the female mice and their offspring were unimmunized.
One week after the second immunization, the female mice were bred with unimmunized male BALB/c mice aged 10 12 weeks in the same cage.
Unimmunized mice were used as a control.
When the basal levels of the various classes of Ig in the sera of unimmunized mice were measured by isotype-specific ELISA, TRAF6-ΔB mice exhibited significantly lower levels of IgM and IgG2b compared to control mice (Fig. 7c).
Sorted Tg T cells from PCC stimulated or unimmunized mice were lysed in TRIzol (Invitrogen, Carlsbad, CA), RNA extracted with the RNeasy kit and genomic DNA removed using the RNase-Free DNase kit (Qiagen, Valencia, CA).
Unimmunized mice were always used as controls (50 µl of VDM alone).
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