Additional file 1 of The therapeutic potential for targeting CSE/H2S signaling in macrophages against Escherichia coli infection

Additional file 1: Key resources table

Additional file 3 of The therapeutic potential for targeting CSE/H2S signaling in macrophages against Escherichia coli infection

Additional file 3: PAG reversed the observed effect on autophagy of CSE overexpression. RAW264.7 cells were transfected with CSE overexpression plasmids, with or without PAG, while control cells were transfected with an empty vector. Immunoblots were performed to detect changes in autophagy-related proteins. Data are presented as the means ± SEMs (n = 3). *(P < 0.05) = significantly different

Additional file 2 of The therapeutic potential for targeting CSE/H2S signaling in macrophages against Escherichia coli infection

Additional file 2: Oligonucleotide sequences used for qPCR

DataSheet_1_Chitosan Protects Immunosuppressed Mice Against Cryptosporidium parvum Infection Through TLR4/STAT1 Signaling Pathways and Gut Microbiota Modulation.docx

Cryptosporidium parvum infection is very common in infants, immunocompromised patients, or in young ruminants, and chitosan supplementation exhibits beneficial effects against the infection caused by C. parvum. This study investigated whether chitosan supplementation modulates the gut microbiota and mediates the TLR4/STAT1 signaling pathways and related cytokines to attenuate C. parvum infection in immunosuppressed mice. Immunosuppressed C57BL/6 mice were divided into five treatment groups. The unchallenged mice received a basal diet (control), and three groups of mice challenged with 1 × 106 C. parvum received a basal diet, a diet supplemented with 50 mg/kg/day paromomycin, and 1 mg/kg/day chitosan, and unchallenged mice treated with 1 mg/kg/day chitosan. Chitosan supplementation regulated serum biochemical indices and significantly (p < 0.01) reduced C. parvum oocyst excretion in infected mice treated with chitosan compared with the infected mice that received no treatment. Chitosan-fed infected mice showed significantly (p < 0.01) decreased mRNA expression levels of interferon-gamma (IFN-γ) and tumor necrosis factor-α (TNF-α) compared to infected mice that received no treatment. Chitosan significantly inhibited TLR4 and upregulated STAT1 protein expression (p < 0.01) in C. parvum-infected mice. 16S rRNA sequencing analysis revealed that chitosan supplementation increased the relative abundance of Bacteroidetes/Bacteroides, while that of Proteobacteria, Tenericutes, Defferribacteres, and Firmicutes decreased (p < 0.05). Overall, the findings revealed that chitosan supplementation can ameliorate C. parvum infection by remodeling the composition of the gut microbiota of mice, leading to mediated STAT1/TLR4 up- and downregulation and decreased production of IFN-γ and TNF-α, and these changes resulted in better resolution and control of C. parvum infection.</p

Comparison of PCR and serum agglutination assays for differentiating the serotypes of APEC isolates and clinical infected samples.

<p>Comparison of PCR and serum agglutination assays for differentiating the serotypes of APEC isolates and clinical infected samples.</p

Sensitivity analysis of the allele-specific PCR assay.

<p>(<b>A</b>) <b>Sensitivity analysis using the bacterial genomic DNA.</b> The detection limit was determined as 10 pg of bacterial DNA for <i>E. coli</i> serotypes O2 and O18 strains, and 500 pg of bacterial DNA for <i>E. coli</i> serotypes O1 and O78 strains, respectively. (<b>B</b>) <b>Sensitivity analysis using the bacterial culture.</b> The detection limit was determined as 10 CFUs of <i>E. coli</i> serotypes O2 and O18 strains, and 1,000 CFUs of <i>E. coli</i> serotypes O1 and O78 strains, respectively. Lane M: DL2000 Marker.</p

Sensitivity analysis of the allele-specific PCR assay.

<p>(<b>A</b>) <b>Sensitivity analysis using the bacterial genomic DNA.</b> The detection limit was determined as 10 pg of bacterial DNA for <i>E. coli</i> serotypes O2 and O18 strains, and 500 pg of bacterial DNA for <i>E. coli</i> serotypes O1 and O78 strains, respectively. (<b>B</b>) <b>Sensitivity analysis using the bacterial culture.</b> The detection limit was determined as 10 CFUs of <i>E. coli</i> serotypes O2 and O18 strains, and 1,000 CFUs of <i>E. coli</i> serotypes O1 and O78 strains, respectively. Lane M: DL2000 Marker.</p

The product profiles of <i>E. coli</i> serotypes O1, O2, O18 and O78 strains amplified using the allele-specific PCR.

<p>Lane M: DL2000 DNA Marker; O1, O2, O18 and O78 represent PCR products for O1, O2, O18 and O78 strains respectively. Lane 1: APEC O1 strain; Lane 2: APEC strain DE47; Lane 3: APEC strain DE14; Lane 4: APEC strain DE17; Lane 5: APEC strain RS218; Lane 6: APEC strain CE66; Lane 7: APEC strain DE48; Lane 8: APEC strain DE65; Lane 9: Negative control.</p

The <i>rfb</i> gene clusters of <i>E. coli</i> serotypes O1, O2, O18 and O78 strains.

<p>The black arrows correspond to <i>gnd</i> and <i>galF</i> genes. Grey arrows correspond to <i>rfb</i> gene cluster and the gene names are <i>italic</i> indicated. The length of <i>rfb</i> gene cluster was also shown. In the PCR reaction system, the universal forward primer was used for all the sero-typing amplification with specific reverse primers. The bold lines below the <i>gnd</i> gene indicate the size of the PCR products for different <i>E. coli</i> serotype strains, which allow the differentiation of the O types. Primers and their locations were also indicated.</p

Primers used in this study.

a<p>The primers were designed based on the gene sequences of <i>wekO</i>, <i>wekS</i>, <i>wekW</i> and <i>wzx</i> in the <i>rfb</i> gene cluster of respective serotypes of <i>E. coli</i> strains.</p