Original images for Western blotting.

(DOCX)</p

Original values used to build graphs.

(XLSX)</p

S1 Fig -

Tlr11 knock-down efficiency in RAW264.7 cells 24 h (A) and 48 h (B) after transfection.Tlr11 expression levels were knocked-down by siRNA targeting Tlr11 (siTlr11). A control siRNA (siCtrl) that targets a scrambled sequence were used as control. 24 h after transfection, RAW264.7 cells were treated with 4 μg/ml MIC3 and OVA for another 24 h. The mRNA levels of Tlr11 and Gapdh were measured 24 h and 48 h after siRNA transfection using qRT-PCR. 1 = no change. Each bar indicates the mean value ± S.D. (n = 3 independent replicates). (TIF)</p

Additional file 2: of Evaluation of factors influencing the guide to read biomedical English literature course for Chinese new medical postgraduates—a multiple regression analysis

Questionnaire for the Literature Guide. (DOCX 25 kb

Protein structure simulation of <i>T</i>. <i>gondii</i> MIC3 peptide.

SWISS-MODEL (https://swissmodel.expasy.org/) was used to simulate the tertiary structure of MIC3 peptide. The model validation parameters are as follows: MolProbity Score 2.54, Clash Score 3.81, Ramachandran Favoured 77.78%. (TIF)</p

Amino acid sequence alignment between <i>T</i>. <i>gondii</i> MIC3 peptide and <i>T</i>. <i>gondii</i> profilin-like protein.

Software Bioedit was used to compare the amino acid sequences of T. gondii MIC3 peptide and profilin-like protein. (TIF)</p

The location of MIC3 in RAW264.7 cells detected by laser confocal microscopy.

To characterize the cellular localization of MIC3, cells were incubated with primary anti-MIC3 mouse monoclonal antibody (Anti-MIC3) and PE-conjugated antibody against mouse F4/80 overnight at 4°C. After being washed, cells were incubated with secondary Alexa Fluor 488 AffiniPure goat anti-mouse IgG antibody (Anti-IgG). DAPI: cell nucleus; F4/80: macrophage surface marker; MIC3: microneme protein 3 of T. gondii.</p

The role of MyD88 in inflammatory immune response induced by MIC3 in RAW264.7 cells.

(A) The levels of TNF-α secreted by RAW264.7 cells after 24 h-treatment of 4 μg/ml MIC3 and OVA. To inhibit the activity of MyD88, RAW264.7 cells in group MIC3+ST2825 were preincubated with 20 μg/ml MyD88 inhibitor ST2825 for 3 h. The secretion of TNF-α was analyzed by ELISA. Each bar indicates the mean value ± S.D. (n = 4 independent replicates). Significance was analyzed using one-way ANOVA. ***, P (B) The mRNA levels of Tnf-α and polarization-related genes after the pretreatment of MyD88 inhibitor ST2825 and 24 h-treatment of MIC3 in RAW264.7 cells. Data are expressed as the means ± S.D. (n = 4 independent replicates). 1 = no change. (C) Ly6C expression in MyD88-inhibited RAW264.7 cells. Ly6C expression in RAW264.7 cells preincubated with 20 μg/ml MyD88 inhibitor ST2825 and treated with 4 μg/ml OVA and 4 μg/ml MIC3 were evaluated by flow cytometry. One representative of three independent experiments is shown. (D) Overlay of representative histograms showing Ly6C expression in MyD88-inhibited RAW264.7 cells treated with 4 μg/ml OVA and 4 μg/ml MIC3. (E) The percentage of Ly6C+ cells in F4/80+CD11b+ gated RAW264.7 cells. Data are expressed as the means ± S.D. (n = 3 independent experiments). Significance was analyzed using one-way ANOVA. ***, P (F) The MFI of Ly6C expression in F4/80+CD11b+ gated RAW264.7 cells. Each bar indicates the mean value ± S.D. (n = 3 independent experiments). Significance was analyzed using one-way ANOVA. ***, P (G) The effects of MyD88 inhibitor ST2825 on MIC3 and LPS-induced NF-κB p65 phosphorylation. The NF-κB p65 phosphorylation of RAW264.7 cells was evaluated by Western blotting.</p

Primers used in this study.

(DOCX)</p

The expression levels of inflammation markers in RAW264.7 cells after 24 h-treatment of MIC3.

(A) Ly6C expression in RAW264.7 cells. After 24 h-treatment of 4 μg/ml OVA and 4 μg/ml MIC3, Ly6C expression on F4/80+CD11b+ gated RAW264.7 cells were evaluated by flow cytometry. One representative of three independent experiments is shown. (B) Overlay of representative histograms showing Ly6C expression in RAW264.7 cells treated with 4 μg/ml OVA and 4 μg/ml MIC3. (C) The percentage of Ly6C+ cells in F4/80+CD11b+ gated RAW264.7 cells. Data are expressed as the means ± S.D. (n = 3 independent experiments). Significance was determined by the two-tailed Student’s t-test. ***, P (D) The mean fluorescence intensity (MFI) of Ly6C expression in F4/80+CD11b+ gated RAW264.7 cells. Data are expressed as the means ± S.D. of three independent samples for each group. Significance was determined by the two-tailed Student’s t-test. ***, P (E) The mRNA levels of inflammatory genes after 24 h-treatment of 4 μg/ml MIC3 in RAW264.7 cells. Data are expressed as the means ± S.D. of four independent samples for each group.</p