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Page 6 of 16 Ding et al. Microbiome Res Rep 2024;3:16 https://dx.doi.org/10.20517/mrr.2023.64

Figure 2. Effects of B. longum subsp. infantis on the levels of cytokines and Immunoglobulin in the colon of neonatal female (A-E) and
male mice (F-J). (A) IFN-γ; (B) IgG2a; (C) IL-4; (D) IgE; (E) the ratio of IgG2a and IgE; (F) IFN-γ; (G) IgG2a; (H) IL-4; (I) IgE; (J) the
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ratio of IgG2a and IgE. P < 0.05, P < 0.01, compared to corresponding gender control group. The mean ± SEM (n = 8 per group) was
used to represent the data. SEM: Error of the mean.
and G]. B. longum subsp. infantis I4MI decreased the level of IL-4 (P < 0.05, Figure 2H), while I8TI and
B6MNI decreased the level of IgE (P < 0.05, Figure 2I) significantly. Furthermore, I4MI, I8TI, and B6MNI
increased the ratio of IgG2a/IgE significantly (P < 0.05, Figure 2J).

Effects of B. longum subsp. infantis on mRNA relative expression of JAK/STAT pathway in the
colon of mice
Based on the described results, B. longum subsp. infantis I4MI, I5MI, I10TI, and B6MNI for female mice,
and I4MI, I5TI, I8TI, and B6MNI for male mice can adjust Th1/Th2 related genes and proteins. Therefore,
these strains were selected for subsequent analysis. The expression of genes associated with the JAK/STAT
pathway in the colon was assessed by qRT-PCR to analyze the effects of B. longum subsp. infantis on their
transcriptional levels. The relative abundance of JAK1 mRNA in the I4MI, I5TI, and B6MNI groups was
significantly higher than that in the female control group (P < 0.05, Figure 3A). No significant differences
were found in the relative expression of JAK2 in the colon of female mice [Figure 3B]. The relative
expression of STAT1 mRNA was significantly higher in I5TI (P < 0.05, respectively) and B6MNI groups
(P < 0.01 for both, Figure 3C) and the relative expression of STAT6 mRNA was significantly lower in I5TI
(P < 0.05) and B6MNI groups (P < 0.01, Figure 3D) compared to that in the female control group. The
relative expression of T-bet mRNA was significantly higher in I5TI (P < 0.05, respectively) and B6MNI
groups (P < 0.01 for both, Figure 3E). No significant differences were found in the relative expression of