Fig 2: CtBP mRNA and protein levels were significantly increased in OA biopsies. (A and B) The relative mRNA levels of CtBP1 and CtBP2. Total RNA from biopsies of 48 OA patients and 48 controls was used to determine the mRNA levels of CtBP1 (A) and CtBP2 (B) by qRT-PCR analyses. The expression of CtBP1 and CtBP2 in a healthy control was defined as one-fold. **P<0.01. (C and D) The protein levels of CtBPs and OA markers in three-paired biopsies. Total cell extracts of three-paired biopsies from OA patients and controls were subjected to western blotting to examine the protein levels of CtBP1, CtBP2, CD31, CD55 and CD68 (C). GAPDH was used as a loading control. (D) Statistical data of the relative protein levels of CtBPs and OA markers in (C). **P<0.01. (E) IHC staining results of CtBPs and OA markers. The same biopsies as (C) were used to IHC staining and representative CtBP1, CtBP2, CD31, CD55 and CD68 images were shown. Bars=100 µm. The positive cell numbers were counted and shown in the right panels. C: Control; O: OA. ***P<0.001.

Fig 3: In vitro and in vivo pro-angiogenic effects of HSD-R. (A) Evaluation of flow velocity and spatial vascular profiles by LASCA. Red polygons show the regions of interest. (B) The quantified mean flux values. (C–D) Representative immunofluorescence images show CD31+ cells (C) and quantitative analysis of the vascularization degree (D). Scale bar, 100 µm. In images of (A to D): Sham, thoracotomy without LAD ligation; MI, LAD ligation plus saline injection; HSD-R (L), LAD ligation and HSD-R injection at 25 µg/kg; HSD-R (H), LAD ligation and HSD-R injection at 100 µg/kg (E–F) Representative images (E) and quantitative analysis (F) of the tube formation by HUVECs. Scale bar, 500 µm. (G–H) Representative images (G) and quantification (H) of migration of HUVECs at various time points after different treatments. Scale bar, 50 µm. (I) RT-qPCR analysis of the mRNA expression levels of typical pro-angiogenesis genes including VEGFa, Fgf1, and Wnt5a. Data are expressed as means ± SD from three independent replicates. For statistical results in (H): ***P < 0.001 versus the control at 12 h; #P < 0.05, ###P < 0.001 versus the control at 24 h. In other cases: *P < 0.05, **P < 0.01, ***P < 0.001; ns, no significance. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig 4: Perivascular cells are slow-cycling.a Left: Distribution of ki67-positive cells in the perivascular and parenchymal compartment in orthotopic S24 (n = 16 regions from three mice, two-tailed t-test) and P3xx (n = 12 regions from four mice, two-tailed t-test) xenograft tumors. Right: ki67 proliferation index (ki67 positive cells/all cells in the respective compartment) of S24 (n = 3 mice, two-tailed t-test) and P3xx tumor cells (n = 4 mice, two-tailed t-test) in the perivascular and the parenchymal compartment in xenograft tumors. b Exemplary immunofluorescence stainings of ki67 (yellow, proliferating cells), nestin (green, tumor cells), DAPI (blue, nuclei), and CD31 (red, endothelium) in S24 and P3xx xenografts. c Left: Quantification of the distribution of EdU-positive tumor cells (indicating cells in S-phase) after in vivo incorporation of EdU (4 h) in the parenchymal and perivascular compartment (S24 xenografts, n = 25 regions from 5 mice, two-tailed t-test; P3xx xenografts, n = 12 regions from 4 mice, two-tailed t-test). Right: EdU labeling index (EdU positive cells/all cells in the respective compartment) in the perivascular and the parenchymal compartment (S24 xenografts, n = 5 mice, two-tailed t-test; P3xx xenografts, n = 4 mice, two-tailed t-test). d Exemplary immunofluorescence staining of EdU (green, cells in S-phase), nestin (red, tumor cells), DAPI (blue, nuclei) and CD31 (yellow, endothelium) demonstrates that most proliferating cells are located in the parenchymal compartment (P3xx xenograft). Blood vessels are marked with arrowheads. e Mitotic index (number of mitotic events normalized to the distribution of cells in the perivascular and parenchymal compartment) of S24 and T269 GBMSCs (n = 16 regions in eight mice (S24), n = 10 regions in 4 mice (T269), two-tailed Mann Whitney tests). Right: Exemplary in vivo 2-PM of mitosis of a parenchymal S24 GBMSC (arrowhead). Cytoplasm (red), nucleus (green), blood vessels (blue). f Distribution of ki67 positive cells in the parenchymal and perivascular compartment in molecularly defined human glioma specimen (oligodendroglioma: n = 16 patients; astrocytoma: n = 19 patients; high grade astrocytoma: n = 15 patients; glioblastoma: n = 16 patients; one-way ANOVA (p = 1.36 × 10-139), Tukey’s post hoc test for multiple comparisons). g Exemplary immunohistochemical stainings of ki67 in molecularly defined human glioma specimen. Arrowheads exemplarily mark blood vessels. Data in violin plots a, c, e, f are represented as median and quartiles. Data in columns a are represented as mean + SEM. *p < 0.05**, p < 0.01, ***p < 0.001. (mut: mutant; LOH: loss of heterozygosity; wt: wild-type). Source data are provided as a Source Data file.

Fig 5: Up-regulation of prothrombin kringle-2 (pKr-2) and neuroinflammatory molecules in the hippocampi of patients with Alzheimer's disease (AD) and five familial Alzheimer's disease (5XFAD) mice. (a) Details of human post-mortem hippocampal tissues presented from Victorian Brain Bank Network. PMI, post-mortem interval. (b) Western blotting for the expression levels of pKr-2, Iba1 and neuronal nuclei (NeuN) in the human hippocampi of age-matched controls (CON) and patients with AD. *P < 0.05 vs. CON (Unpaired Student's t-test; n = 5 for each group). (c) Western blotting for the protein levels of TLR4, TNF-a, IL-1ß and iNOS in the human hippocampi of age-matched controls (CON) and patients with AD. *P < 0.05, versus CON (Unpaired Student's t-test; n = 5 for each group). (d) Western blotting for the protein levels of pKr-2, Iba1 and NeuN in the hippocampal tissues obtained from 3-, 6- and 9-month-old 5XFAD and 9-month-old WT mice. *P < 0.05 versus 9-month-old WT mice (one-way ANOVA with Tukey's post hoc analysis; n = 5 for each group). (e) Western blotting for the protein levels of TLR4, TNF-a, IL-1ß and iNOS in the hippocampal tissues obtained from 5XFAD and WT mice. *P < 0.05 and versus 9-month-old WT mice (one-way ANOVA with Tukey's post hoc analysis; n = 5 for each group). All quantitative values are presented as the mean ± SEM. (f) Location of pKr-2 expression in the hippocampi of 5XFAD mice. The red fluorescent signals represent Iba1, GFAP, NeuN and CD31, and the green fluorescent signal represents pKr-2, respectively, in the respective hippocampal section of 9-month-old WT and 5XFAD mice. The white arrowheads indicate pKr-2 expression. Scale bar, 50 µm