Additional file 1 of Helicobacter pylori outer membrane vesicles induce astrocyte reactivity through nuclear factor-κappa B activation and cause neuronal damage in vivo in a murine model
Creators: name: Palacios, Esteban
name: Lobos-González, Lorena
name: Guerrero, Simón
name: Kogan, Marcelo J.
name: Shao, Baohai
name: Heinecke, Jay W.
name: Quest, Andrew F. G.
name: Leyton, Lisette
name: Valenzuela-Valderrama, Manuel
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Neuroscience
subject: Immunology
subject: FOS: Clinical medicine
subject: 111714 Mental Health
subject: FOS: Health sciences
subject: 110309 Infectious Diseases

description: Additional file 1. Table S1. Identification of proteins with unique peptides in Helicobacter pylori (Hp) 60190 OMVs. The results of the LC–MS/MS analysis and the search using the Hp UniProtKB database (UP000000429.fasta) allowed to identify the total protein content of OMVs and the parental bacteria Hp 60190. The table shows the protein ID and the average number of unique peptides of proteins found in OMVs. Figure S1. Evaluation of the dose-response of the reactivity markers in DITNC1 ATCC astrocytes treated with different concentrations of OMVs from Helicobacter pylori (Hp) 60190. DITNC1 ATCC cells were incubated in the absence or presence of OMVs (1.25 to 20 µg/ml) from Hp 60190, at 37 °C for 24 h. As a positive reactivity control, the astrocytes were incubated with TNF (10 ng/ml, 48 h). After treatment, whole cell lysates were evaluated by immunoblot analysis of total connexin 43 (A), β3 integrin (B), GFAP (C), and vimentin (D), normalized to β-actin. Values in the graphs were obtained by averaging the immune-specific band intensity normalized to β-actin from 3 independent experiments (mean ± S.E.M). *p &lt; 0.05 and **p &lt; 0.01, compared to controls (Ctrl). Figure S2. Dose-response of the reactivity markers in DITNC1 ATCC astrocytes treated with 2.5 µg/ml of Helicobacter pylori (Hp) OMVs at different times. DITNC1 ATCC cells were incubated in the absence or presence of 2.5 µg/ml of Hp 60190 OMVs at 37 °C, from 3 to 72 h. As a positive reactivity control, the astrocytes were incubated with TNF (10 ng/ml, 48 h). After treatment, whole cell lysates were evaluated by immunoblot analysis of total connexin 43 (A), β3 integrin (B), GFAP (C), and vimentin (D), normalized to β-actin. Values in the graphs were obtained by averaging the immune-specific band intensity, normalized to β-actin, from 3 independent experiments (mean ± S.E.M). *p &lt; 0,05, **p &lt; 0.01, and ***p &lt; 0.001, compared to controls (Ctrl). Figure S3. Effect of Helicobacter pylori (Hp) OMVs on astrocyte cell viability. Primary astrocytes (post 17 days in vitro) were incubated in the absence or presence of 2.5 µg/ml of OMVs for 12 h. Cell viability was evaluated with the Trypan blue assay in a Neubauer chamber. Values in the graph represent the average of the percentage of negative trypan blue cells from 3 independent experiments (mean ± S.E.M.). Figure S4. Effect of Helicobacter pylori (Hp) OMVs on p65 NF-ĸB subunit protein levels in DITNC1 ATCC astrocytes, following different times of exposure. DITNC1 ATCC cells were incubated in the absence or presence of 2.5 µg/ml of OMVs from 3 to 72 h, at 37 °C. As a positive reactivity control, the astrocytes were incubated with TNF (10 ng/ml, 48 h). After treatment, whole cell lysates were evaluated by immunoblot analysis of p65 NF-ĸB total levels. Values in the graph were obtained by averaging the immune-specific band intensity, normalized to β-actin, from 3 independent experiments (mean ± S.E.M). *p &lt; 0.05, **p &lt; 0.01, and ***p &lt; 0.001, compared to controls (Ctrl). Table S2. Research Resource IdentifiersList of reagents, antibodies, inhibitors, kits, experimental models used in this study. Tee catalog number is included when available.