Journal: Journal of Hematology & Oncology

Article Title: Challenges and advances in clinical applications of mesenchymal stromal cells

doi: 10.1186/s13045-021-01037-x

Figure Lengend Snippet: The tests of modified MSCs using CRISPR-Cas9 technology

Article Snippet: Immortalized human bone marrow MSC cell line (ATCC PCS-500–041) , PUMILIO2 (PUM2) , Depletion of PUM2 blocks MSC adipogenesis and enhances osteogenesis. PUM2 works as a negative regulator on the 3′ UTRs of JAK2 and RUNX2 via direct binding. CRISPR/CAS9-mediated gene silencing of Pum2 inhibited lipid accumulation and excessive bone formation , [ ] .

Techniques: Modification, CRISPR, Functional Assay, Synthesized, In Vitro, Transfection, Expressing, Transplantation Assay, Inhibition, Binding Assay

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Colorimetric staining and quantification of ALP activity in hBM-MSCs cultured for seven days on collagen type I-coated (a) and fibronectin-coated (b) substrates. Results are expressed as mean ± SD (n=3). Data were analyzed assuming a Gaussian (normal) distribution and equal variances across groups; statistical differences were assessed using one-way ANOVA. Only p-values <0.1 are shown. Scale bar represents 200 µm.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Staining, Activity Assay, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Fluorescence images of hBM-MSCs cultured on β-PVDF films coated with either collagen type I (a) or fibronectin (b) and corresponding morphological features. Nuclei are stained in blue, vinculin in green and F-actin in red. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown. Scale bars represent 200 µm.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Fluorescence, Cell Culture, Staining

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: MYPT1 phosphorylation in hBM-MSCs cultured on collagen type I-coated (a) and fibronectin-coated (b) β-PVDF films of varying surface potential. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Phospho-proteomics, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Fluorescence images of hBM-MSCs cultured on β-PVDF films coated with either collagen type I (a) or fibronectin (b) immunostained against YAP and counterstained with Hoechst 33342, and the corresponding quantifications (right panels). Scale bars represent 200 µm. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Fluorescence, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Volcano plots with the −log 10 (p-value) plotted against their respective log 2 (fold change) of genes differentially expressed in hBM-MSCs, Venn diagrams and histogram plots showing genes that are down- or upregulated (p<0.05) in hBM-MSCs cultured on the indicated surfaces for 24 hours (a) or four days (b). Circle area in a and b is proportional to the number of genes. GSEA of the cells cultured on the different surfaces for 24 hours (c) and 4 days (d).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: (a) Immunostaining of hBM-MSCs cultured on the different β-PVDF surfaces with glutaraldehyde-crosslinked collagen type I coating and corresponding morphologic analysis (b). MYTP1 phosphorylation (c) and YAP translocation (d and e) in hBM-MSCs cultured on the substrates. Only p-values <0.1 are shown. Scale bars represent 200 µm. Results are expressed as mean ± SD (n=3).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Immunostaining, Cell Culture, Phospho-proteomics, Translocation Assay

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Immunofluorescent images (a) and corresponding morphologic analysis (b) of hBM-MSCs treated with the ROCK inhibitor Y-27632 for 24 hours. YAP immunolocalization (c) and translocation (d) in treated cells. Only p-values <0.1 are shown. Scale bars represent 300 µm. Results are expressed as mean ± SD (n=4).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Translocation Assay

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: C5a promoted arterial thrombosis by triggering neutrophils to release NETs. A , B H&E staining of thrombus, cross-sections ( A ) and longitudinal sections ( B ). Scale bar = 100 µm. C Quantification of the thrombus size in cross-sections by H&E staining ( n = 5 each). D Thrombus weight ( n = 5 each). E The blood flow velocity in the LICA was dramatically decreased in the FeCl 3 -induced thrombus mice compared with the sham mice, and this change was reversed by PMX53. F Quantification of the blood flow velocity ( n = 4 each). G Immunofluorescence staining of cross-sections of the LCCA for CitH3 (red), Ly6g (green) and DAPI (blue) 6 h after FeCl 3 exposure. Thirty minutes before FeCl 3 exposure, PMX53 was administered by intraperitoneal injection. Scale bar = 75 µm. H Quantification of NET formation capacity shown as the ratio of NETs/neutrophil in LCCA cross-sections subjected to immunofluorescence staining (NaCl group = 3, PMX53 group = 4). I Representative images of immunofluorescence stainings of NET formation for DNA (SYTOX green), CitH3 (red), Ly6g(blue) in vitro after stimulation of C5a, and NET formation was attenuated by PMX53. Scale bar = 75 µm. J Quantification of NET formation capacity in vitro shown as the percentage of NET release, which was assessed by immunofluorescence staining ( n = 3 each). (K) NETosis was measured using a plate reader assay. NET release is expressed as percentage of total DNA ( n = 3 each). Data are presented as mean ± SD

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: Staining, Immunofluorescence, Injection, In Vitro

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: Interactions among C5a, mitochondrial STAT3 and NETs. A , B Western blot analysis was performed to test the expression levels of mitochondrial STAT3 and p-STAT3 (Ser 727 ) in neutrophil-like cells cocultured with C5a, compared with the control. VDAC, a marker of mitochondria, was used as a loading control for mitochondria ( n = 5 each). C NET release in response to buffer or AG490 was measured using a plate reader assay ( n = 3 each). D Representative images of immunofluorescence staining for DNA (SYTOX green), CitH3 (red) and Ly6g (blue) in vitro after stimulation with AG490 showing the presence of NETs. Scale bar = 100 µm. E Quantification of NET formation capacity shown as the percentage of NET release in vitro after stimulation with buffer or AG490, as assessed by immunofluorescence staining. ( n = 3 each). Data are presented as mean ± SD

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Control, Marker, Immunofluorescence, Staining, In Vitro

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: The reduction in arterial thrombotic burden induced by PMX53 was abolished by AG490 in vivo. A , B H&E staining of thrombus, cross-sections A and longitudinal sections B . The thrombus area was reduced by PMX53, and AG490 abolished this effect. A Scale bar = 100 µm; B Scale bar = 200 µm. C Quantification of the thrombus size in cross-sections by H&E staining ( n = 5 each). D Thrombus weight ( n = 5 each). E Blood flow velocity in the LICA. AG490 reversed the increased in blood flow induced by PMX53. F Quantification of the blood flow velocity ( n = 4 each). G Immunofluorescence staining of cross-sections of the LCCA for CitH3 (red), Ly6g (green) and DAPI (blue) 6 h after FeCl 3 exposure. Scale bar = 75 µm. H Quantification of NET formation capacity shown as the ratio of NETs/neutrophil in LCCA cross-sections subjected to immunofluorescence staining (NaCl group = 3; PMX53 group = 4; PMX53 + AG490 group = 3; AG490 group = 4). Data are presented as mean ± SD

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: In Vivo, Staining, Immunofluorescence

Journal: Thrombosis Journal

Article Title: Complement C5a induces the generation of neutrophil extracellular traps by inhibiting mitochondrial STAT3 to promote the development of arterial thrombosis

doi: 10.1186/s12959-022-00384-0

Figure Lengend Snippet: Visual summary: the effect of complement C5a in arterial thrombosis. In arterial thrombosis, C5a chemotactically attracts neutrophils to migrate towards the culprit site and triggers the release of NETs, which contribute to thrombosis by promoting coagulation and stabilizing clots. C5a-induced promotion of NET release is dependent on Mito-ROS production. C5a induces the Mito-ROS production by inhibiting mitochondrial STAT3 activity

Article Snippet: Mouse neutrophils were isolated from the bone marrow of tibias and femurs from healthy C57BL/6 J mice using the Neutrophil Isolation Kit (Solarbio, China) following the manufacturer’s instructions.

Techniques: Coagulation, Activity Assay

Journal: Kidney Diseases

Article Title: Clinical and Experimental Insights into the Role of NETosis in IgA Nephropathy Pathogenesis

doi: 10.1159/000546343

Figure Lengend Snippet: Elevated serum NETosis markers in IgAN patients correlate with CitH3 and Gd-IgA1 levels, and increased NETosis markers in renal tissue. a–c Comparison of serum levels of Citrullinated Histone H3 (CitH3), myeloperoxidase (MPO), and neutrophil elastase (NE) between IgAN patients ( n = 16) and healthy controls (HC, n = 13). d–f Pearson correlation analysis between serum CitH3, MPO, NE, and Gd-IgA1 levels. g Immunofluorescence staining of CitH3 and MPO in kidney tissues from IgAN ( n = 5) and minimal change disease (MCD, n = 5) patients. h–i Quantitative analysis of immunofluorescence staining intensity in the kidneys. Unpaired t tests were used for group comparisons, and Pearson correlation was employed for correlation analysis. Data are presented as mean ± SEM.

Article Snippet: Bone marrow was harvested from healthy C57BL/6J mice, and neutrophils were isolated utilizing a mouse bone marrow neutrophil isolation kit (Solarbio, P8550).

Techniques: Comparison, Immunofluorescence, Staining

Journal: Molecular medicine reports

Article Title: Long non‑coding RNA SNHG14 affects the proliferation and apoptosis of childhood acute myeloid leukaemia cells by modulating the miR‑193b‑3p/MCL1 axis.

doi: 10.3892/mmr.2020.11729

Figure Lengend Snippet: Figure 1. SNHG14 gene expression is upregulated in bone marrow tissues of patients with AML and AML cell lines. (A) Relative expression of SNHG14 in 57 AML bone marrow tissues and NBM. (B) Relative expression of SNHG14 in AML cell lines and human normal bone marrow CD34+ cells. **P<0.01 vs. CD34+ cells. The 2‑ΔΔCq method was used to analyse the relative mRNA expression level using GAPDH as an internal control. AML, acute myeloid leukaemia; NBM, normal marrow tissues; SNHG14, small nucleolar RNA host gene 14.

Article Snippet: Human normal bone marrow CD34+ cells and AML cell lines (MV‐4‐11, AML‐193, HL‐60, and KG‐1 cells) were obtained from the American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C.

Techniques: Gene Expression, Expressing, Control

Journal: Molecular medicine reports

Article Title: Long non‑coding RNA SNHG14 affects the proliferation and apoptosis of childhood acute myeloid leukaemia cells by modulating the miR‑193b‑3p/MCL1 axis.

doi: 10.3892/mmr.2020.11729

Figure Lengend Snippet: Figure 3. miR‑193b‑3p is a target of SNHG14 in AML cells. (A) Starbase was used to predict the binding site between SNHG14 and miR‑193b‑3p. (B) Relative expression of miR‑193b‑3p in MV‑4‑11 and AML‑193 cells following SNHG14 silencing. **P<0.01 vs. blank control. (C) The target association between SNHG14 and miR‑193b‑3p was determined using an RNA immunoprecipitation assay. **P<0.01 vs. Anti‑IgG. (D) The target association between SNHG14 and miR‑193b‑3p was determined using a dual luciferase reporter gene assay. **P<0.01 vs. miR‑NC. (E) Relative expression of miR‑193b‑3p in 57 AML bone marrow tissues and NBM was detected by RT‑qPCR. (F) Spearman's correlation analysis was performed to evaluate the correlation between SNHG14 and miR‑193b‑3p expression. (G) Relative expression of miR‑193b‑3p in AML cell lines and human normal bone marrow CD34+ cells. **P<0.01 vs. CD34+ cells. The 2‑ΔΔCq method was used to analyse the relative mRNA expression level using U6 as an internal control. SNHG14, small nucleolar RNA host gene 14; miR, microRNA; si, small interfering RNA; NC, negative control; wt, wildtype; mut, mutated; NBM, normal marrow tissues; AGO2, protein argonaute‑2; AML, acute myeloid leukaemia.

Article Snippet: Human normal bone marrow CD34+ cells and AML cell lines (MV‐4‐11, AML‐193, HL‐60, and KG‐1 cells) were obtained from the American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C.

Techniques: Binding Assay, Expressing, Control, RNA Immunoprecipitation, Luciferase, Reporter Gene Assay, Small Interfering RNA, Negative Control

Journal: Molecular medicine reports

Article Title: Long non‑coding RNA SNHG14 affects the proliferation and apoptosis of childhood acute myeloid leukaemia cells by modulating the miR‑193b‑3p/MCL1 axis.

doi: 10.3892/mmr.2020.11729

Figure Lengend Snippet: Figure 5. miR‑193b‑3p targets MCL1 in AML cells. (A) TargetScan was used to predict the binding site between miR‑193b‑3p and MCL1. (B) A dual luciferase reporter gene assay was employed to verify the target association between miR‑193b‑3p and MCL1. **P<0.01 vs. miR‑NC. (C) Relative expression of MCL1 in 57 AML bone marrow tissues and NBM was detected by RT‑qPCR. (D) Spearman's correlation analysis was performed to evaluate the correlation between MCL1 and miR‑193b‑3p expression. (E) Spearman's correlation analysis was performed to evaluate the correlation between MCL1 and SNHG14 expression. (F) Relative expression of miR‑193b‑3p in AML cell lines and human normal bone marrow CD34+ cells. **P<0.01 vs. CD34+ cells. The 2‑ΔΔCq method was used to analyse the relative mRNA expression level using GAPDH as an internal control. SNHG14, small nucleolar RNA host gene 14; miR, microRNA; NC, negative control; wt, wildtype; mut, mutated; NBM, normal marrow tissues; AML, acute myeloid leukaemia; MCL1, MCL1 apoptosis regulator BCL2 family member.

Article Snippet: Human normal bone marrow CD34+ cells and AML cell lines (MV‐4‐11, AML‐193, HL‐60, and KG‐1 cells) were obtained from the American Type Culture Collection and cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone; GE Healthcare) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C.

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Expressing, Control, Negative Control