Fig 2: No significant accumulation of M2 marophages in P7 PAB groups hearts. (A) FACS gating scheme for the identification of CD45+CD11b+F4/80+Ly6c– M2 macrophage. (B) Quantification of M2 macrophage as the cell number per mg of tissue (n = 3). Data are presented as Mean ± SEM, Statistic analysis was performed with two-tailed Student’s t-test. n.s., non-significant.

Fig 3: Immunofluorescence staining for retinal vasculature (lectin, green) and macrophages (F4/80, red) of retinas from mice with OIR and age-matched normal controls at different time points. The eyes of mice with OIR and age-matched control mice were fixed and retinas were dissected, incubated in FITC-isolectin B4 and PE-labeled anti-mouse F4/80 for 45 min at room temperature. Retinas were then flat-mounted and examined by fluorescence microscopy. Ischemic retinas from mice with OIR exhibited small tufts and bulbous networks of neovascularization from the retinal capillary bed (A–F) from P12 to P24, and a close association with macrophages was observed by counterstaining with F4/80 [(H–M) and (A'–F') merged images]. Clear vasculature of normal mice was examined (N–S) counterstained with F4/80 [(T–Y) and (N'–S') merged images] (n=4 mice/group). OIR, oxygen-induced retinopathy; NOR, normal; P, post-natal day.

Fig 4: Hepatocytes and Kupffer cells purified from the mouse liver(A) The morphology of hepatocytes two days after plating.(B) The morphology of Kupffer cells. In both (A) and (B), the scale bar is 20 µm.(C–F) Analysis of the purity of Kupffer cells by flow cytometry. Kupffer cells on petri dishes were collected by trypsinization at 37°C for five minutes, and 2 × 105 Kupffer cells were incubated with isotype control antibodies (red) or antibodies against cell markers of interest (blue) on ice for 30 min. Cells were then rinsed with DPBS three times and subjected to flow cytometry for the analysis of F4/80 (C), dead cells using Fixable Blue Dead Cell Staining Kit (D), CD11b and CD45 (E), and TIM-4 and CD45 (F). Panel (C) was adapted from Figure S1A of Li et al. (Li et al., 2022). Panels (D) and (E) were from different experiments.

Fig 5: Loxl1 knockout mice recapitulate the molecular changes in vagina tissue of clinical POP. (A) Heat map across all the samples using the top 500 most differently expressed genes in vagina tissue between the Loxl1 knockout mice and WT mice. (B–D) gene ontology categories of differently expressed genes, horizontal axis represent the fold enrichment, p value < 0.05. (E, F) Venn diagram (left) and the form (right) showed the overlaps of up-regulated (E) and down-regulated (F) differentially expressed genes enriched GO terms between human vagina (H-vagina) and mice vagina (M-vagina). (G, H) Immunofluorescence staining for visualization (red) of immune cell infiltration in human vagina (G) and mice vagina (H). The number of positive cells (red) in each High Power Field of vision (HPF) were quantified respectively. (CD45: leukocyte, CD68: macrophage in human, F4/80: macrophage in mice; White dotted line indicated the interface between mucus layer and submucosa layer, n = 5 in each group).