Fig 2: T. gondii infection down-regulates Trem2 expression on decidual macrophages in pregnant mice.(A) Representative image of the placentas and fetuses of wildtype mice at G 17.5 infected with T. gondii. Red arrow indicated fetal demise. (B and C) Fetal development assessed by fetal size (CRL×OF) and fetal weight. Each data points represent individual fetuses (n = 5–7 mice). CRL: crown-rump length; OF: occipito-frontal diameter. (D) Representative HE staining showing pathological features of mouse placentas. The normal mouse placental structure is composed of decidua zone (De), junctional zone (JZ), and labyrinth zone (LZ). Pathological necrosis in wildtype mice infected with T. gondii mainly occurs in the De. (E) Representative flow cytogram of CD11b+ F4/80+ decidual macrophages, comparing decidual macrophage proportions in normal or infected placentas in mice. The placentas of each mouse were divided into three groups for technically repeated experiments. Data point represents the placenta of a single pregnant mouse (n = 7–8 mice). (F) Representative MFI of Trem2 on the surface of CD11b+ F4/80+ decidual macrophages in normal or infected placentas in mice. The placentas of each mouse were divided into three groups for technically repeated experiments. Data point represents the placenta of a single pregnant mouse (n = 7–8 mice). MFI: mean fluorescence intensity. (G) Decidual macrophages were labeled with F4/80, and the colocalization between Trem2 (red) and F4/80 (green) was imaged by immunohistofluorescence. (H) Immunohistochemistry of normal and infected mouse placentas, which was immunostained with anti-Trem2 and counterstained with hematoxylin. (I) Immunoblot of Trem2 expression and statistical analysis on normal and infected mouse placentas. Each data point represents the placenta of an individual pregnant mouse (n = 8 mice). Data were presented as mean ± SD. Statistical analysis was conducted using two-tailed unpaired Student’s t-test (B, C, E, F and I). *: P < 0.05.

Fig 3: Confocal immunofluorescence of integrins and inflammatory response proteins with/without Ti treatment. (A-C) Localization of CD18, CD11b and CD11a after Ti co-culture. (D and E) Confocal images of Myd88 and TIRAP localizations. Non-linear adjustments. (F) Statistical analysis was performed among multiple marks comparing fluorescence ratio. Green represents anti-IgA staining. Scale bar, 10 µm. *P<0.05. Ti, titanium; TIRAP, TIR-domain-containing adapter protein.

Fig 4: Western blotting and quantitative analysis of integrin levels and inflammation response proteins. (A) Western blot immunoblots results showing CD18, CD11a, CD11b, CD11c and MyD88 TIRAP. (B-D) Quantification of immunoblots using ImageJ software. *P<0.05. Ti, titanium; TIRAP, TIR-domain-containing adapter protein.

Fig 5: PARVB deficiency protects against cisplatin-induced renal inflammation. (A) Representative images of kidney sections stained with antibody against CD11b (a myeloid cell marker) in WT and PARVB cKO mice after cisplatin (15 mg/kg) or vehicle treatment. Scale bar: 50 μm. Quantification analysis was shown in the right panel. ***P < 0.001 vs. WT + Cisplatin. n = 3 mice for each group. (B) Representative images of kidney sections stained with antibody against F4/80 (a macrophage marker) in WT and PARVB cKO mice after cisplatin (15 mg/kg) or vehicle treatment. Scale bar: 20 μm. Quantification analysis was shown in the right panel. ***P < 0.001 vs. WT + Cisplatin. n = 6 mice for each group. (C) ELISA analysis of TNF-ɑ level in mouse serum isolated from WT or PARVB cKO mice with or without cisplatin (15 mg/kg) treatment. **P < 0.01 vs. WT + Cisplatin. n = 5 mice for each group. (D-E) qPCR analysis of TNF-ɑ(D) and IL-1β(E) mRNA expression in control (Nc siRNA) and PARVB knockdown (PB siRNA1 and PB siRNA2) HK2 cells with or without cisplatin (15 µM) treatment. *P < 0.05, **P < 0.01 vs. HK2 + Cisplatin, n = 3 independent experiments