We used qPCR to assess the integrity of two circulating miRNAs in mouse serum, serum treated with EDTA, and plasma collected with EDTA.
In vitro, anti-mouse mesangial cell serum treated mesangial cells showed greater release of lactate dehydrogenase, decreased cell survival, and increased apoptotic cell death.
B. pertussis-immune serum with a titer of 6400, a titer similar to that of B. bronchiseptica-induced immune serum, reduced B. bronchiseptica numbers in both organs by>99.9% relative to that of naïve serum treated mice, an effect similar to that of B. bronchiseptica-immune serum (Figure 3A).
At 6 days after infection, the reduction (approximately 110-fold) of HRSV replication in the lungs of SHe-KLH immune serum treated mice was more pronounced than at 4 days post-infection (Supplementary Fig S7C).
In TLR4-deficient mice, however, the co-inoculation had no effect on B. parapertussis numbers in either naïve serum treated or immune serum treated mice (Fig. 7A).
After washing with PBST (3×), the mouse serum-treated ubiquitin peptide conjugates were added to each well and incubated for 2 h.
In short, mouse serum samples were treated at room temperature for 1 h and Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Invitrogen) was used as the detection antibody.
A marginal, but significant serum pH difference (p < 0.03) was observed between DB and DCA treated mouse serum.
Briefly, 25 µl of mice serum was first treated with 50 µl of receptor destroying enzyme II (catalog number YCC 340 122; Accurate Chemical and Scientific, Westbury NY) at a 1∶3 ratio (vol/vol) at 37°C overnight.
Mice treated with 10 µl or 20 µl antibodies displayed both significantly fewer CFU compared to the mice treated with normal mouse serum (Table 4, experiments 1 and 2).
