Journal: bioRxiv

Article Title: Major histocompatibility complex class II-expressing bone marrow megakaryocytes activate CD4 + T cells and induce regulatory T cell fate

doi: 10.1101/2025.11.21.689743

Figure Lengend Snippet: (A) Representative images (left) and quantification of MK numbers and MK area (right) in MHC KO and Cre ( - ) mice; n =10 mice/ genotype. (B) Colocalization analysis of LAP and CD41 on femoral cryosections of MHC KO and Cre ( - ) mice; n =4 mice/ genotype. (C) Total MK counts 24 hours after LPS or Poly (I:C) administration; n= 5 mice/ genotype. (D) Frequency of CD40, CD53, CD80, CD83, CD86 (+) MKs 24 hours after LPS or Poly (I:C) administration; n = 5 mice/ genotype. Data are shown as mean ± SD; graphs represent data from at least 3 independent experiments. Statistics performed with unpaired 2-tailed Student t test (A–C) and 1-way ANOVA with Tukey’ multiple comparisons test at 95.00% CI (C).

Article Snippet: MKs and platelets were stained overnight using antibodies against LAP (AF246-NA, R&D Systems), a-tubulin (322588, ThermoFisher Scientific), proplatelet basic protein (PPBP), (PA5-47947, Invitrogen), CD42a (M-051-0, Emfret Analytics), Ovalbumin, Alexa FluorTM 647 Conjugate (Thermo Fisher Scientific), nuclei were stained using DAPI.

Techniques:

Journal: bioRxiv

Article Title: Cardiomyocyte-expressed TGFβ signals to fibroblasts to program early heart maturation and adult myocyte identity

doi: 10.1101/2025.09.22.677845

Figure Lengend Snippet: a . Western blot showing time course of TGFβ1 or Latency associated peptide (LAP) (TGFβ) expression in the heart at embryonic day 16.5 (E16.5), postnatal day 1 (P1), P7, P21, and 12 weeks of age. Gapdh was used to normalize protein loading. b . Western blot for TGFβ1 and Cre from protein extracts of neonatal cardiomyocytes from Tgfb1/2/3 fl/fl-αMHC-Cre mice transduced with Ad-GFP or Ad-Cre 48 hrs prior in culture. c . mRNA expression of Tgfb1 , Tgfb2 , and Tgfb3 from neonatal cardiomyocytes transduced with Ad-GFP (adenovirus expressing green fluorescent protein) or Ad-Cre (adenovirus expressing Cre recombinase). n=2-5 per treatment group, *p<0.05. d . Schematic of the generation of Tgfb1/2/3 fl/fl-αMHC-Cre mice whereby Tgfb1 fl/fl , Tgfb2 fl/fl , and Tgfb3 fl/fl mice were crossed to the αMHC-Cre transgene. e . Kaplan-Meier curve depicting survival of Tgfb1/2/3 fl/fl and Tgfb1/2/3 fl/fl-αMHC-Cre mice over time in weeks. f . Heart-weight to body-weight (HW/BW) ratio of Tgfb1/2/3 fl/fl , Tgfb1/2/3 fl/fl-αMHC-Cre , and αMHC-Cre mice at 6 weeks of age. n=4-19 mice in each group, *p<0.05. g . Fractional shortening (FS%) at 6 weeks of age of the same groups of mice as in f. n=6-17 mice per group. *p<0.05. h . Left ventricular internal diameter in diastole (LVID;d) at 6 weeks of age in the same groups of mice as in f. n=6-17 mice per group. *p<0.05. i . Representative histological sections from hearts stained with Sirius Red to assess collagen deposition and wheat germ agglutinin-FITC (WGA, green) to assess cardiomyocyte cross sectional area (CSA) of Tgfb1/2/3 fl/fl and Tgfb1/2/3 fl/fl-αMHC-Cre mice at 6 weeks of age. Scale bar Sirius Red: 2 mm; Scale bar WGA: 100 µm. j . Quantification of average cardiomyocyte CSA from WGA staining as shown in i. A minimum of 250 myocytes were measured from each heart, across 3 hearts (points on the graph). *p<0.05. k . Hydroxyproline quantification of fibrosis from the left ventricle of hearts at 6 weeks of age as shown in j. n=8-9 mice per group. *p<0.05. l . Flow cytometry quantification of MEFSK4+ fibroblasts at 6 weeks of age in the hearts of mice shown in j. n=9-10 mice per group. *p<0.05. c,j,l. Data are presented as mean +/-SEM. Statistical analysis was performed using Student’s T-tests. f-h. Statistical analysis was performed using One Way ANOVAs with Tukey’s multiple comparisons test. k. In the case of non-normally distributed data, Mann Whitney tests were performed. Data are presented as mean +/-SEM.

Article Snippet: Antibodies against the following proteins were used: periostin (Novus Biologicals, Centennial, CO, USA; NBP1-30042; 1:100 dilution for IF, 1:500 dilution for Western blot); Collagen I (Abcam, Boston, MA, USA; ab21286; 1:100 for IF, 1:500 for Western blot), TGFβ1 (Abcam 215715; 1:500 for Western blot), LAP (TGFβ1) (R&D systems, AF-246-NA, Minneapolis, MN; 1:200 for Western Blot), Gapdh (Fitzgerald, #10R-G109A for Western Blot), phospho-SMAD2 (Ser465/467)/phospho-SMAD3 (Ser423/425) (Cell Signaling Technology, #8828, Danvers, MA; 1:500 for Western blot), phospho-SMAD3 (Ser423/425) (Cell Signaling Technology, #9520; 1:500 for Western blot), SMAD3 (Abcam #ab40854; 1:500 for Western blot), SMAD2 (Cell Signaling Technology, # 5339; 1:500 for Western blot), αSMA (Sigma-Aldrich #A2547; 1:100 for IHC), lumican (Abcam #168348; 1:500 for Western Blot), transglutaminase 2 (TGM2, Abcam, #2386; 1:500 for Western Blot), pHH3(Cell Signaling, 9701; 1:100), desmin (Abcam, #15200; 1:100 for IHC), connexin 43 (Cx43, Cell Signaling, 3512; 1:100 for IF), laminin (Sigma-Aldrich, #LO663; 1:100 for IF), SERCA1 (Thermo Scientific, MA3-912, 1:100 for IF), fibronectin (FN, Abcam, #2413; 1:500 for Western Blot), fibrillin-2 (Santa Cruz Biotechnology, Santa Cruz, CA; sc-393968), loxl2 (Invitrogen, PA5-85210), pleiotrophin (Thermo Scientific, PA5-94984), tenascin C (Abcam, #6346), WT1 (Abcam, Ab89901), Cre (EMD Millipore, 69050-3).

Techniques: Western Blot, Expressing, Transduction, Staining, Flow Cytometry, MANN-WHITNEY

Journal: bioRxiv

Article Title: Cardiomyocyte-expressed TGFβ signals to fibroblasts to program early heart maturation and adult myocyte identity

doi: 10.1101/2025.09.22.677845

Figure Lengend Snippet: a . Uniform Manifold Approximation and Projections (UMAPs) displaying cellularity from hearts of Tgfb1/2/3 fl/fl and Tgfb1/2/3 fl/fl-αMHC-Cre mice at P7. b . Composite scoring of genes associated with embryonic cardiomyocytes based on expression of Myh7 , Tnni1 , Myl4 , and Myl7 . Each dot represents a nucleus scored based on enrichment in the embryonic cardiomyocyte gene signature. c . Composite scoring of relative expression of mature cardiomyocyte gene expression assessed by expression of Myh6 , Tnni3 , Myl3 , and Myl2 . Each dot represents a nucleus scored based on enrichment in the embryonic cardiomyocyte gene signature. d . Composite scoring of relative gene expression associated with mitochondrial maturation-based genes listed in Supplementary Table 3. e . Violin plot showing relative expression of Lamb1 , Lamb2 , Lamc1 , and Itgb1 in cardiomyocytes from Tgfb1/2/3 fl/fl (red) and Tgfb1/2/3 fl/fl-αMHC-Cre (blue). f . CellChat analysis of the number of inferred interactions and interaction strength between cardiac fibroblast ligands (cell-cell interactions or ECM-cell interactions) and cardiomyocyte receptors from hearts of Tgfb1/2/3 fl/fl-αMHC-Cre mice and littermate controls. g . Dotplot of predicted differential ligand and receptor interactions between fibroblast (Fib) ligands paired with cardiomyocyte (CM) receptors. The scale located at the top of the figure displays strength of the predicted interaction. h . Schematic model of embryonic heart development where TGFβ generated by cardiomyocytes signals to tissue resident cardiac fibroblasts to initiate their differentiation and generation of proper ECM, matricellular, and growth factors (GF) that collectively signaling to maturing cardiomyocytes to ensure their differentiation and identity.

Article Snippet: Antibodies against the following proteins were used: periostin (Novus Biologicals, Centennial, CO, USA; NBP1-30042; 1:100 dilution for IF, 1:500 dilution for Western blot); Collagen I (Abcam, Boston, MA, USA; ab21286; 1:100 for IF, 1:500 for Western blot), TGFβ1 (Abcam 215715; 1:500 for Western blot), LAP (TGFβ1) (R&D systems, AF-246-NA, Minneapolis, MN; 1:200 for Western Blot), Gapdh (Fitzgerald, #10R-G109A for Western Blot), phospho-SMAD2 (Ser465/467)/phospho-SMAD3 (Ser423/425) (Cell Signaling Technology, #8828, Danvers, MA; 1:500 for Western blot), phospho-SMAD3 (Ser423/425) (Cell Signaling Technology, #9520; 1:500 for Western blot), SMAD3 (Abcam #ab40854; 1:500 for Western blot), SMAD2 (Cell Signaling Technology, # 5339; 1:500 for Western blot), αSMA (Sigma-Aldrich #A2547; 1:100 for IHC), lumican (Abcam #168348; 1:500 for Western Blot), transglutaminase 2 (TGM2, Abcam, #2386; 1:500 for Western Blot), pHH3(Cell Signaling, 9701; 1:100), desmin (Abcam, #15200; 1:100 for IHC), connexin 43 (Cx43, Cell Signaling, 3512; 1:100 for IF), laminin (Sigma-Aldrich, #LO663; 1:100 for IF), SERCA1 (Thermo Scientific, MA3-912, 1:100 for IF), fibronectin (FN, Abcam, #2413; 1:500 for Western Blot), fibrillin-2 (Santa Cruz Biotechnology, Santa Cruz, CA; sc-393968), loxl2 (Invitrogen, PA5-85210), pleiotrophin (Thermo Scientific, PA5-94984), tenascin C (Abcam, #6346), WT1 (Abcam, Ab89901), Cre (EMD Millipore, 69050-3).

Techniques: Expressing, Gene Expression, Generated

Journal: bioRxiv

Article Title: Inhibition of RhoA-mediated secretory autophagy in megakaryocytes mitigates myelofibrosis in mice

doi: 10.1101/2024.12.04.626665

Figure Lengend Snippet: (A) Analysis of transforming growth factor β1 (TGFβ1) levels in bone marrow fluid derived from C57BL/6N, thrombopoietin (TPO)-deficient ( Tpo −/- ) or MPL-deficient mice ( Mpl −/- ). n=6-10 mice. One-way ANOVA with Dunnett’s test for multiple comparisons. (B) Visualization of latency-associated peptide (LAP)/TGFβ1, von Willebrand Factor (vWF) and the megakaryocyte (MK) marker CD42b in femoral cryosections. Nuclei were counterstained using DAPI. Scale bars: 20 µm. (C, D) Correlation analysis between CD42b and vWF (C) and CD42b and LAP/TGFβ1 (D) . Each dot represents one MK. n=2 mice. (E, F) Analysis of TGFβ1 and PF4 levels in cell culture supernatant after incubation of HSPCs in the absence (Control) or presence of TPO. Values were normalized to TPO values. n=8 mice. Unpaired, two-tailed Student’s t-test. (G) Visualization of LAP/TGFβ1, vWF and proplatelet basic protein (PPBP) in round and proplatelet-forming MKs. Goat (gt) and rabbit (rb) isotype antibodies served as staining controls. Nuclei were counterstained using DAPI. Scale bars: left: 10 µm; right: 5 µm. (H) Pearson colocalization coefficient analysis using Costes threshold of proplatelet-forming MKs imaged using super-resolution microscopy (Airyscan Leica LSM880, 63x objective). Each dot represents one MK. LAP/TGFβ1 served as a positive control. (I) Visualization of LC3B, LAP/TGFβ1 and CD42b in native bone marrow MKs. Nuclei were counterstained using DAPI. Scale bars: 15 µm. (J) Pearson colocalization coefficient analysis using Costes threshold for LC3B and LAP/TGFβ1. Each dot represents one MK. n=2 mice. (K) Correlation analysis between LAP/TGFβ1 and LC3B in cultured, bone marrow-derived MKs. Each dot represents one MK. n=3 mice. (L, M) Visualization of vWF and LAP/TGFβ1 (L) as well as LC3B and LAP/TGFβ1 (M) in platelets adhered on a fibrinogen-coated surface. Platelets were stained for α-tubulin to highlight the cytoskeleton. Scale bars: 5 µm. All data are presented as mean ± SD.

Article Snippet: Sections were rehydrated in PBS for 20 min, non-specific binding was blocked using 10% donkey serum and sections were stained for either CD105 (AF1320, R&D Systems), laminin (L9393, Sigma-Aldrich), LAP (AF-246-NA, R&D Systems), von Willebrand factor (vWF) (PA5-16634, ThermoFisher Scientific), collagen I (ab21286, abcam), collagen IV (ab6586, abcam), CD41 (133902, BioLegend) or CD42b (M-051-0, Emfret Analytics).

Techniques: Derivative Assay, Marker, Cell Culture, Incubation, Control, Two Tailed Test, Staining, Super-Resolution Microscopy, Positive Control

Journal: bioRxiv

Article Title: Inhibition of RhoA-mediated secretory autophagy in megakaryocytes mitigates myelofibrosis in mice

doi: 10.1101/2024.12.04.626665

Figure Lengend Snippet: (A) White blood cells (WBC) count of mice transplanted with CD117 + MSCV-EGFP (Control)- or MSCV-IRES-EGFP-MPL W515L (MPL W515L )-transduced cells three weeks after transplantation. n=4 mice. Unpaired, two-tailed Student’s t-test. (B-D) Platelet count (B) , mean platelet volume (MPV) (C) and immature platelet fraction (IPF) (D) of mice transplanted with CD117 + Control or MPL W515L -transduced cells. n=4 mice. Unpaired, two-tailed Student’s t-test. (E, F) Percentage of EGFP + platelets (E) and CD45 + cells (F) of mice transplanted with Control or MPL W515L -transduced cells three weeks after transplantation. n=4. Unpaired, two-tailed Student’s t-test. (G) Spleen to body weight of mice transplanted with Control or MPL W515L -transduced cells. n=4 mice. Unpaired, two-tailed Student’s t-test. (H-J) Visualization (H) and quantification of MFI of CD41 (I) and TGFβ1/LAP (J) in femoral cryosections of mice transplanted with Control or MPL W515L -transduced cells. At least 50 MKs were analyzed per mouse. n=3 mice. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD. (K-M) Visualization (K) and quantification of collagen I (L) and collagen IV deposition (M) in femoral cryosections of mice transplanted with Control or MPL W515L -transduced cells. n=3 mice. Each dot represents one field of view (FOV). Three FOVs were analyzed per mouse. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD. (N, O) Analysis of TGFβ1 (N) and PF4 levels (O) in cell culture supernatant of CD117 + Control or MPL W515L -transduced cells after culture in the presence of TPO for 72h. n=4 mice. Unpaired, two-tailed Student’s t-test. (P, Q) LC3B immunoblot (P) and densitometric analysis (Q) of platelets derived from mice transplanted with CD117 + Control or MPL W515L -transduced cells. n=6 mice. Unpaired, two-tailed Student’s t-test. (R, S) RhoA immunoblot (R) and densitometric analysis (S) of platelets derived from mice transplanted with CD117 + Control or MPL W515L -transduced cells. n=6 mice. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD.

Article Snippet: Sections were rehydrated in PBS for 20 min, non-specific binding was blocked using 10% donkey serum and sections were stained for either CD105 (AF1320, R&D Systems), laminin (L9393, Sigma-Aldrich), LAP (AF-246-NA, R&D Systems), von Willebrand factor (vWF) (PA5-16634, ThermoFisher Scientific), collagen I (ab21286, abcam), collagen IV (ab6586, abcam), CD41 (133902, BioLegend) or CD42b (M-051-0, Emfret Analytics).

Techniques: Control, Transplantation Assay, Two Tailed Test, Cell Culture, Western Blot, Derivative Assay

Journal: bioRxiv

Article Title: Inhibition of RhoA-mediated secretory autophagy in megakaryocytes mitigates myelofibrosis in mice

doi: 10.1101/2024.12.04.626665

Figure Lengend Snippet: (A) Schematic of autophagy-related gene 5 (ATG5) in autophagosome formation. (B) Schematic of the MPL W515L transplant model in mice. (C) Transduction efficiency of CD117 + cells derived from WT and Atg5 HSC-KO mice transduced with MPL W515L one or four days after transduction. n=4. Multiple, unpaired Student’s t-tests. (D, E) Percentage of EGFP + CD45 + cells (D) and platelets (E) of WT and Atg5 HSC-KO mice transplanted with Control or MPL W515L -transduced cells four weeks after transplantation. n=5-6 mice. Two-way ANOVA with Sidak’s correction for multiple comparisons. (F) Spleen to body weight of WT and Atg5 HSC-KO mice transplanted with Control or MPL W515L -transduced cells. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. (G, H) White blood cell (WBC) (G) and red blood cell (RBC) counts (H) of WT and Atg5 HSC-KO mice. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. (I, J) Platelet counts (I) and immature platelet fraction (IPF) (J) of WT and Atg5 HSC-KO mice. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. (K, L) Visualization (K) and quantification (L) of splenic MKs of transplanted WT and Atg5 HSC-KO mice. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. Scale bars: 500 µm. (M) Quantification of MKs and megakaryocyte progenitors (MkP) in the bone marrow of MPL W515L mice transplanted with WT or Atg5 HSC-KO cells by flow cytometry. n=5-6 mice. Two-way ANOVA with Sidak’s correction for multiple comparisons. (N) Quantification of MK numbers in femoral cryosections of MPL W515L mice transplanted with WT or Atg5 HSC-KO cells. n=5 mice. Unpaired, two-tailed Student’s t-test. (O-Q) Visualization (O) and quantification of intracellular LAP/TGFβ1 (P) and collagen I (Q) in MPL W515L mice transplanted with WT or Atg5 HSC-KO cells. n=5-6 mice. Unpaired, two-tailed Student’s t-test. Scale bars: 50 µm. (R-T) Analysis of TGFβ1 (R) , IL1β (S) and PF4 levels (T ) in bone marrow fluid of MPL W515L mice transplanted with WT or Atg5 HSC-KO cells. n=5-6 mice. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD. Schematics were generated using Biorender.com.

Article Snippet: Sections were rehydrated in PBS for 20 min, non-specific binding was blocked using 10% donkey serum and sections were stained for either CD105 (AF1320, R&D Systems), laminin (L9393, Sigma-Aldrich), LAP (AF-246-NA, R&D Systems), von Willebrand factor (vWF) (PA5-16634, ThermoFisher Scientific), collagen I (ab21286, abcam), collagen IV (ab6586, abcam), CD41 (133902, BioLegend) or CD42b (M-051-0, Emfret Analytics).

Techniques: Transduction, Derivative Assay, Control, Transplantation Assay, Flow Cytometry, Two Tailed Test, Generated

Journal: bioRxiv

Article Title: Inhibition of RhoA-mediated secretory autophagy in megakaryocytes mitigates myelofibrosis in mice

doi: 10.1101/2024.12.04.626665

Figure Lengend Snippet: (A) Transduction efficiency of CD117 + cells derived from WT and RhoA MK-KO mice transduced with MPL W515L one day after transduction. n=3 mice. Multiple unpaired Student’s t-tests. (B, C) Percentage of EGFP + CD45 + cells (B) and platelets (C) of WT and RhoA MK-KO mice transplanted with Control or MPL W515L -transduced cells four weeks after transplantation. n=5-6 mice. Two-way ANOVA with Sidak’s correction for multiple comparisons and unpaired, two-tailed Student’s t-test. (D) Spleen to body weight of WT and RhoA MK-KO mice transplanted with Control or MPL W515L -transduced cells. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. (E, F) White blood cell (WBC) (E) and red blood cell (RBC) counts (F) of transplanted WT and RhoA MK-KO mice. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. (G) Platelet counts of transplanted WT and RhoA MK-KO mice. n=5-6 mice. One-way ANOVA with Sidak’s correction for multiple comparisons. (H, I) Visualization (H) and quantification (I) of splenic MKs of MPL W515L -transplanted WT and RhoA MK-KO mice. n=6 mice. Scale bars: 500 µm. Unpaired, two-tailed Student’s t-test. (J) Quantification of MK numbers in femoral cryosections of WT and RhoA MK-KO mice transplanted with MPL W515L -transduced cells. n=6 mice. Unpaired, two-tailed Student’s t-test. (K-M) Visualization (K) and quantification of collagen IV (L) and intracellular LAP/TGFβ1 (M) and in transplanted WT and RhoA MK-KO mice. n=6. Unpaired, two-tailed Student’s t-test. Scale bars: 50 µm. (N-P) Analysis of TGFβ1 (N) , IL1β (O) and PF4 levels (P ) in bone marrow fluid of MPL W515L -transplanted WT and RhoA MK-KO mice. n=6 mice. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD.

Article Snippet: Sections were rehydrated in PBS for 20 min, non-specific binding was blocked using 10% donkey serum and sections were stained for either CD105 (AF1320, R&D Systems), laminin (L9393, Sigma-Aldrich), LAP (AF-246-NA, R&D Systems), von Willebrand factor (vWF) (PA5-16634, ThermoFisher Scientific), collagen I (ab21286, abcam), collagen IV (ab6586, abcam), CD41 (133902, BioLegend) or CD42b (M-051-0, Emfret Analytics).

Techniques: Transduction, Derivative Assay, Control, Transplantation Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Inhibition of RhoA-mediated secretory autophagy in megakaryocytes mitigates myelofibrosis in mice

doi: 10.1101/2024.12.04.626665

Figure Lengend Snippet: (A, B) Percentage of EGFP + CD45 + cells (A) and platelets (B) of PBS- or Y27632 (Y)-treated mice transplanted with MPL W515L -transduced cells three weeks after transplantation. n=5 mice. Two-way ANOVA with Sidak’s correction for multiple comparisons and unpaired, two-tailed Student’s t-test. (C) Spleen to body weight of transplanted PBS- and Y-treated mice. n=5 mice. Unpaired, two-tailed Student’s t-test. (D, E) White blood cell (WBC) (D) and red blood cell (RBC) counts (E) of transplanted PBS- and Y-treated mice. n=5 mice. Unpaired, two-tailed Student’s t-test. (F-H) Platelet count (F), mean platelet volume (MPV) (G) and immature platelet fraction (IPF) (H) in transplanted PBS- and HQ-treated mice. n=5 mice. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD. (I-K) Visualization (I) and quantification of intracellular LAP/TGFβ1 (J) and collagen IV (K) in transplanted PBS- and Y-treated mice. n=5. Unpaired, two-tailed Student’s t-test. Scale bars: 50 µm. (L) Quantification of MK numbers in femoral cryosections in transplanted PBS- and Y-treated mice. n=5 mice. Unpaired, two-tailed Student’s t-test. (M-O) Analysis of TGFβ1 (M) , IL1β (N) and PF4 levels (O ) in bone marrow fluid of transplanted PBS- and Y-treated mice. n=5 mice. Unpaired, two-tailed Student’s t-test. All data are presented as mean ± SD. Schematic was generated using Biorender.com.

Article Snippet: Sections were rehydrated in PBS for 20 min, non-specific binding was blocked using 10% donkey serum and sections were stained for either CD105 (AF1320, R&D Systems), laminin (L9393, Sigma-Aldrich), LAP (AF-246-NA, R&D Systems), von Willebrand factor (vWF) (PA5-16634, ThermoFisher Scientific), collagen I (ab21286, abcam), collagen IV (ab6586, abcam), CD41 (133902, BioLegend) or CD42b (M-051-0, Emfret Analytics).

Techniques: Transplantation Assay, Two Tailed Test, Generated