Additional file 1 of Bacteriocin Microcin J25’s antibacterial infection effects and novel non-microbial regulatory mechanisms: differential regulation of dopaminergic receptors
Creators: name: Shang, Lijun
name: Yang, Fengjuan
name: Chen, Qingyun
name: Dai, Ziqi
name: Yang, Guangxin
name: Zeng, Xiangfang
name: Qiao, Shiyan
name: Yu, Haitao
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Information Systems not elsewhere classified
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Computational Biology

description: Additional file 1: Table S1 Ingredient composition of the experimental diets. Table S2 Primers used in the study. Table S3 Scoring system for histological changes in the jejunum. Table S4 Components of the used volume densities. Table S5 Primary and secondary antibodies used for immunofluorescence and western blot labeling. Table S6 Environmental factor screening based on VIF analysis. Fig. S1 F4 susceptibility screening results. (A) For infection model establishment. (B) For J25/Gen treatment. Fig. S2 Effects of graded levels ETEC challenge on piglets. (A) Experimental design. (B) Fecal fluid content analysis. n = 18, mean ± SEM; *, compared with the NC group, P &lt; 0.05. (C) Diarrhea score: 0, normal; 1, loose stool; 2, moderate diarrhea; 3, severe diarrhea. n = 18, mean ± SEM. (D) Cytokines measured in the serum on d 0, after ETEC challenge for 24 h, and on d 5. n = 6, mean ± SEM. (E) Total bacteria and E. coli populations in the feces. n = 6, mean ± SEM. ANOVA followed by Tukey’s multiple comparisons test; different lowercase letters within each group indicate significantly different values (P &lt; 0.05). Fig. S3 J25’s effects on jejunum morphology, inflammation status and fecal microbiota output. (A) Morphological measurement of the jejunum. (B–C) Cytokines in the serum. (D) Cytokines in the jejunum. (E) Fecal E.coli and total bacteria detected on d 0, ETEC 24 h and d 5 by PCR. n = 8, mean ± SEM. ANOVA followed by Tukey’s multiple comparisons test; different lowercase letters within each group indicate significantly different values (P &lt; 0.05). Fig. S4 J25’s effects on ETEC-challenged mice. (A) Experimental design of repeated trials in the mouse model. (B) Body weight changes (relative to original weight, set as 100%), n = 18, mean ± SEM. (C) Diarrhea score. Scoring standards: 0, normal; 1, loose stool; 2, moderate diarrhea; 3, severe diarrhea. n = 18, mean ± SEM. (D) Representative images of the jejunum by H&amp;E staining and histopathological scores. n = 6, mean ± SEM. (E) Cytokines levels in the serum. n = 6, mean ± SEM. (F and G) Representative FACS plots of CD3−CD49b+ (F) and F4/80+ CD11b+ (G) staining of cells isolated from mouse jejunums, and quantification of the results. n = 4, mean ± SEM. ANOVA followed by Tukey’s multiple comparisons test; different lowercase letters within each group indicate significantly different values (P &lt; 0.05). Fig. S5 Relative abundance of predicted metabolic potential of microbes from ETEC and ETEC + J25 groups, as predicted by PICRUSt2. Fig. S6 Correlation matrix between microbiota and important parameters. Only genera with statistically significantly change are shown. Spearman correlation, *, P &lt; 0.05; **, P &lt; 0.01; ***, P &lt; 0.001. Fig. S7 Significantly altered metabolites in neuron-related pathways identified by the metabolome. (A–F) Metabolites in the dopaminergic neuron pathway. (C, E–H) Metabolites in the serotonergic neuron pathway. (C, D, I, J) Metabolites in the glutamatergic neuron pathway. (I, J) Metabolites in the GABAergic neuron pathway. (C, K, L) Metabolites in the cholinergic neuron pathway. n = 8, mean ± SEM. ANOVA followed by Dunnett’s multiple comparisons test; different lowercase letters within each group indicate significantly different values (P &lt; 0.05). Fig. S8 Effects of graded levels of 6-OHDA lesions on mice. (A) Body weight changes (relative to original weight, set as 100%), d 1, n = 12; d 2–7, n = 6, mean ± SEM. (B) Representative images of TH immunoreactivity by the immunohistochemistry method in the jejunum on d 1 (first, top of B1) and d 6 (second, bottom of B1), and relative immunolabeling quantification (B2). (C) Representative images of the jejunum by H&amp;E staining on d 6 and histopathological scores. n = 6, mean ± SEM. ANOVA followed by Tukey’s multiple comparisons test; different lowercase letters within each group indicate significantly different values (P &lt; 0.05). Fig. S9 Gentamicin-induced resistance was significantly associated with altered microbiota structure. (A) Annotation results of differential metabolites between ETEC and Gen groups. (B) Differential metabolites in the vancomycin resistance pathway. n = 8, mean ± SEM. ANOVA followed by Dunnett’s multiple comparisons test; different lowercase letters within each group indicate significantly different values (P &lt; 0.05). (C) Canonical Correspondence Analysis plot. The arrow length represents the strength of the correlation between the environmental variables and the microbes. The longer the arrow, the stronger the correlation. The perpendicular distance between the microbes and the environmental variable axes on the plot reflects their correlations. The less the distance, the stronger the correlation.