Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: Leptin upregulates MTA1 expression in human breast cancer cells. MTA1 mRNA expression was analyzed using qPCR in MDA-MB-231 ( A ) and Hs 578T cells ( B ) after leptin treatment for 24 h. Results are expressed as mean ± SD; * p < 0.05 and ** p < 0.01 vs. untreated control. MTA1 protein expression was assessed using Western blot in MDA-MB-231 ( C ) and Hs 578T cells ( D ) after 24 h of leptin treatment.

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Expressing, Control, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: Leptin upregulates MTA1 expression via the Ob-R/STAT3 pathway in human breast cancer cells. MTA1 protein expression was evaluated using Western blot in MDA-MB-231 ( A , C ) and Hs 578T cells ( B , D ) after 24 h of leptin treatment in the presence or absence of Ob-R BP ( A , B ) or AG490 ( C , D ).

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: MTA1 overexpression promotes VM in human breast cancer cells. MDA-MB-231 ( A , C , E ) and Hs 578T cells ( B , D , F ) were transfected with the MTA1 CRISPR activation plasmid for 48 h. MTA1 protein expression was assessed using Western blot in MDA-MB-231 ( A ) and Hs 578T cells ( B ). VM was performed using a 3D culture assay for 16 h in MDA-MB-231 ( C ) and Hs 578T cells ( D ) (40× magnification; scale bar = 500 μm). Results are expressed as mean ± SD; *** p < 0.001 vs. control plasmid. Expression of VM-related proteins was evaluated using Western blot in MDA-MB-231 ( E ) and Hs 578T cells ( F ).

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Over Expression, Transfection, CRISPR, Activation Assay, Plasmid Preparation, Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Leptin-Upregulated Metastasis-Associated Protein 1 Promotes Vasculogenic Mimicry in Breast Cancer Cells

doi: 10.3390/ijms26125726

Figure Lengend Snippet: MTA1 silencing inhibits leptin-induced VM in human breast cancer cells. MDA-MB-231 ( A , C , E ) and Hs 578T cells ( B , D , F ) were treated with leptin 48 h after transfection with MTA1 siRNA. MTA1 protein expression was evaluated using Western blot in MDA-MB-231 ( A ) and Hs 578T cells ( B ). VM was performed using a 3D culture assay for 16 h in MDA-MB-231 ( C ) and Hs 578T cells ( D ) (40× magnification; scale bar = 250 μm). Results are expressed as mean ± SD; *** p < 0.001 vs. untreated control; ### p < 0.001 vs. control siRNA. Expression of VM-related proteins was evaluated using Western blot in MDA-MB-231 ( E ) and Hs 578T cells ( F ).

Article Snippet: MDA-MB-231 cells (1.5 × 10 5 ) and Hs 578T cells (2.0 × 10 5 ) were cultured under optimal conditions and transfected with 1 μg of the control or 0.5 μg of the MTA1 CRISPR activation plasmid (Santa Cruz, Danvers, MA, USA) for 48 h. Transfection was performed using the UltraCruz transfection reagent (Santa Cruz), according to the manufacturer’s protocol.

Techniques: Transfection, Expressing, Western Blot, Control

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET deficiency leads to replicative senescence aHDFs (3 × 10 5 cells/mL) were seeded in each culture plate and were treated, in triplicate, without (UT) or treated (T) with C35 (5 μM), peptides (20 μM), TET1 siRNA ( TET1 i ; 300 nM). Cells treated with Bleomycin (Bleo; 5 μg/mL), or H 2 O 2 (100 μM) were used as positive control. Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Heatmap of TET gene expression. (B) TET activity and global DNA levels of 5 mC, 5hmC, and 5 fC. (C) Expression patterns of energy/stress and nutrient-sensing pathways in aging PBMCs mirror those observed in young, actively replicating aHDFs (week 3) undergoing senescence, as well as those treated with TET1 i , C35, and RLS. (D) Expression level of hTERT , NAMPT , and PCNA . (E) Expression level of COL1A1 and ELN. (F) Cellular UPS activity, ATP, NAD + /NADH ratio, and LC3B levels. (G) Cellular levels of LC3B without (−) and after pre-treatment with (+) Bafilomycin A1 (1 μM) for 24 h. (H) Cellular ROS. (I) Extent of oxidative DNA damage assessed by the cellular level of 8OHdG. Positive control was comprised of cells treated with H 2 O 2 and Bleomycin (Bleo). Negative control was comprised of cells treated with CLV. (J) γH2AX immunolocalized to nuclei. For positive control, cells were treated with Bleomycin (Bleo). Intranuclear γH2AX (green) appears as focal spots (red arrows) or as a diffuse pan-nuclear pattern (yellow arrows) in nuclei marked by DAPI (blue) staining. Nuclei are further highlighted by double hashed lines. The boxes in the left pane (scale bars 10 μM) are magnified in the right insets (scale bars 2.5 μm). (K) NFKB1 gene expression. (L) Level of intra-nuclear phosphorylated p65 (p65P). (M) Cellular levels of ROS, IL6, and IL8 were secreted into culture media. (N) Expression levels of senescence markers ( CDKN2A , CDKN1A , CDKN1B , LTA4H , TIMP1 , and MMP1 ). (O) Expression level of LDH toxicity, SAβ-Gal activity, and extracellular level of lactic acid (LA). (P) LDHA1 gene expression. (Q) Rate of proliferation assessed by the quantitation of BrdU incorporated into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. Points are shown as empty and mean points as filled circles. The distribution of data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Positive Control, Incubation, Gene Expression, Activity Assay, Expressing, Negative Control, Staining, Quantitation Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET dependent resilience to damage and protection against damage-induced senescence Proliferating aHDFs (0.3 × 10 6 /mL) and PBMCs (70 years; PBMC 70Yr, 1 × 10 6 /mL) were seeded in culture plates and pre-treated in triplicate without (untreated: UT) or with H 2 O 2 (100 μM) for 24 h. After 24 h, cells were washed and treated without or with CLV (20 μM), or CRISPR TET1 ( TET1 CR ;1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression levels of TETs in PBMCs. (B) AKG bioavailability in PBMCs. (C) Expression levels of energy/stress and nutritional sensors in PBMCs. (D) Heatmap of senescence marker gene expression in PBMCs. (E) Level of extracellular lactic acid (LA) and SAβ-Gal activity in PBMCs. (F) Quantification of BrdU and ROS (qROS) in PBMCs. Bar and line graphs show the means ± SD. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 . Points are shown as empty and mean points as filled circles.

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: CRISPR, Incubation, Expressing, Marker, Gene Expression, Activity Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: AKG-TET deficient senescence state is reversible Young replicatively proliferating aHDFs (week 3, 0.3 × 10 6 /mL)) were seeded in culture plates and pre-treated in triplicates without (UT) and with C35 (5 μM), RLS (20 μM), or TET1 siRNA ( TET1 i ; 300 nM). Aliquot of cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. The remaining cells were re-seeded in equal numbers and cultured without treatment for an additional 7 days (withdrawal). Cultures were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media. (A) Quantification of TET activity, and 5 mC and 5fc levels. (B) Expression level of energy/stress and nutritional sensor genes. (C) Expression level of NFKB1 , RELA , IKBA , COL1A1, and ELN . (D) γH2AX immunolocalized (cyan) in nuclei. Nuclei stained for DAPI (deep blue) are marked by double hashed lines. (E) Extracellular level of lactic acid (LA), SAβ-Gal activity, LDH toxicity, and UPS. (F) Intracellular level of ROS and 8OHdG. (G) Heatmap of senescence markers and RRM2 gene expression. (H) Rate of proliferation assessed by BrdU incorporation into the nuclei of cells in S-phase. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Incubation, Cell Culture, Activity Assay, Expressing, Staining, Gene Expression, BrdU Incorporation Assay

Journal: iScience

Article Title: AKG-TET axis is central to senescence plasticity

doi: 10.1016/j.isci.2025.114298

Figure Lengend Snippet: Activation of the AKG-TET axis reverses replicative and age induced senescence Replicatively induced senescence. Replicatively senescent aHDFs (0.3 × 10 6 cells/mL) were seeded in triplicate in each plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids (TET1 CR , 1 μg/mL). Cells were incubated with 30 μM BrdU for 18 h prior to removal of both the cells and culture media following 7 days of treatment. (A) Expression level of TET s. (B) AKG bioavailability and TET activity (TET act ). (C) Expression level of energy/stress and nutritional sensor gene expression. (D) Intracellular level of ROS. (E) Heatmap of senescence marker and RRM2 gene expression. (F) Level of lactic acid (LA) in culture media. (G) Cellular SAβ-Gal activity. (H) Quantitation of nuclear BrdU. (I) Total cell numbers. Age induced senescence. PBMCs (PBMC 70Yr ) were seeded (1 × 10 6 cells/mL) in triplicate in each well of a 24-well plate and were treated without (UT) and with RLS (20 μM), CLV (20 μM), or CRISPR TET1 plasmids ( TET1 CR : 1 μg/mL). Cells and culture media were removed on day 7 of treatment for analysis. (J) Quantitation of bioavailable AKG and TET activity in cells treated with RLS versus CLV. (K) qPCR quantitation of the expression level of TET s. (L) qPCR quantitation of the expression level of energy/stress and nutritional sensor gene expression. (M) Quantitation of BrdU incorporated into nuclei of cells in S-phase, lactic (LA) acid in culture media, cellular SAβ-Gal activity, and ROS. (N) Heatmap of senescence markers and RRM2 in PBMCs. Bar and line graphs show the means ± SD. Boxplots show the first and third quartiles and median values. All points are shown as empty, and mean points as filled circles. The distribution of all data points is shown by Beeswarm in Violin plots. Statistical significance was assessed using Student’s t test for two-group comparisons. p -values are presented as follows: ns (not significant), p ≤ 5 × 10 −1 , ∗p ≤ 5 × 10 −2 , ∗∗p ≤ 5 × 10 −3 , ∗∗∗p ≤ 5 × 10 −4 , ∗∗∗∗p ≤ 5 × 10 −5 .

Article Snippet: CRISPR activation system for TET1 ( TET1 CR ) , Santa Cruz Biotechnology (Santa Cruz, CA) , sc-400845-ACT-2.

Techniques: Activation Assay, CRISPR, Incubation, Expressing, Activity Assay, Gene Expression, Marker, Quantitation Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 3 WNT11 is a novel ligand for ROR2 in humans. a RNA-Seq of MCF-7 pROR2 cells: Network of differentially expressed genes associated with non-canonical WNT signaling grouped according to their cellular localization. b MCF-7 cells were stimulated for 24 h with rWNT5A (100 ng/ml) and WNT11 expression was analyzed by western Blot. c+d Expression of the non-canonical WNT ligands was measured by qRT-PCR in MCF-7 (c) or the indicated ROR2-overexpressing human breast cancer cell lines (d) (mean ± SD, n = 3–9, *p < 0.05, p < 0.01, n.e. = not expressed). Expression values were calculated relative to the empty vector control cells. e Co-immunoprecipitation (Co-IP) of V5-WNT11 in MCF-7 pROR2 cells detects ROR2 by western blot. f Schematic representation of the ROR2 N-terminal deletion constructs. g Co-IP of V5-Wnt11 in MCF-7 expressing either pROR2-FL or pROR2-ΔΔ. h Cell invasion assays of MCF-7 expressing N-terminal ROR2 deletion constructs (mean ± SD, n = 3, *p < 0.01, **p = 0.0001, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: RNA Sequencing, Expressing, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Control, Immunoprecipitation, Co-Immunoprecipitation Assay, Construct, Comparison

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 4 WNT11 mediates the pro-tumoral effects of ROR2. a+b Invasion assay: MCF-7 pcDNA or pROR2 cells were treated with WNT inhibitors (a) or Porcupine inhibitors (b) (mean ± SD, n = 3, *p < 0.05, **p < 0.01, ***p < 0.0001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c Invasion assay of MCF-7 pROR2 cells stably expressing a non-sense control (ns ctl) or WNT11 (shWNT11) shRNA (mean ± SD, n = 3, *p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. d Invasion of MCF-7 pROR2 cells transfected with control (siCTL) or WNT11 siRNA (siWNT11) +/− rhWNT11 (100 ng/ml) was assessed in Boyden chambers (mean ± SD, n = 3, *p < 0.05, **p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. e Invasion assay of BT-474 pcDNA and pROR2 cells transfected with either siCTL or siWNT11 (mean ± SD, n = 3, *p < 0.05, **p < 0.001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. f AFM of cell-cell-junctions in the indicated cell lines. g Immunofluorescence for the tight junction protein ZO-1. h ECIS measurements of MCF-7 cells (box: 25-75th percentile, line at median, *p < 0.0001). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. i+j xCELLigence measurements of MCF-7 cells (mean ± SD). Shown is one representative example (i) and a corresponding Area Under the Curve (AUC) analysis for all three independent experiments (mean ± SD, *p < 0.01). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: Invasion Assay, Comparison, Stable Transfection, Expressing, Control, shRNA, Transfection, Immunofluorescence

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 5 WNT11 mediates ROR2 signaling. a+b Western blot: RHOA and ROCK2 in MCF-7 and BT-474 pROR2 cells treated with control siRNA (siCTL) or siRNA against WNT11 (siWNT11) (a) with corresponding densitometric quantification normalized on HSP90 expression (b) (mean ± SD, *p < 0.05, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c Levels of active RHOA-GTP in the indicated cells were assessed by ELISA (mean ± SD, *p < 0.05, ns = not significant). Significance was calculated with a one-way ANOVA with Dunnett’s multiple comparison test. c MCF-7 pcDNA and pROR2 siCTL/siWNT11 cells were characterized by RPPA for phospho-proteins associated with the WNT signaling pathway (n = 3, *p < 0.05). d Schematic representation of the master regulator analysis of ROR2/WNT11 signaling. The illustration was created with BioRender.com

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: Western Blot, Control, Expressing, Comparison, Enzyme-linked Immunosorbent Assay

Journal: Journal of experimental & clinical cancer research : CR

Article Title: WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

doi: 10.1186/s13046-021-02187-z

Figure Lengend Snippet: Fig. 6 ROR2/WNT11 are expressed in metastatic breast cancer and associated with poor survival. a Pathway (pw) enrichment for RNA-Seq data from 31 patients with brain metastases given for different WNT subpathways. Significance was calculated with a log rank test. b qRT-PCR: Expression of ROR2 and WNT11 in samples of human brain metastases (line at median). The upper numbers indicate the number of samples with positive signals out of all investigated samples. c Kaplan-Meier survival curves showing the OS of metastatic patients based on their averaged WNT11 and ROR2 expression. The separation high/low was computed based on an optimal cutoff using the maxstat method. Significance was calculated with a log rank test

Article Snippet: The vector for V5-tagged active WNT11 was a gift from Xi He (Addgene plasmid #43824; http:// n2t. net/ addge ne: 43824; RRID:Addgene_43,824) [16].

Techniques: RNA Sequencing, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Wnt5a-mediated Adipo-Cardiac Interorgan Communication in HFpEF

doi: 10.1101/2025.10.29.685456

Figure Lengend Snippet: (A) Schematic outline of study. (B) Cellular localization of differentially expressed genes (DEGs) in Nprc -AKO visceral adipose tissue (VAT). (C) Increased and decreased gene ontologies in Nprc -AKO vs control VAT. (D) Volcano plot of DEGs in Nprc -AKO vs control VAT (red indicates p < 0.01 and 50%-fold-change compared to control). (E) Reverse transcriptase quantitative PCR of top 5 Wnt isoforms in Nprc -AKO vs control VAT. (F) Plasma circulating levels of Wnt5a in global (GKO), cardiomyocyte-specific (CKO), adipocyte-specific (AKO), and control (WT) mice after L-NAME and high fat diet. (G) Cell size in H9C2 cells after treatment with Wnt5a. (H) RT-qPCR for hypertrophic markers NPPA, NPPB, and MYH7 in H9C2 cells after Wnt5a treatment.

Article Snippet: For studies investigating the effects of Wnt5a treatment, cells were transfected with either pcDNA3.2-Wnt5a plasmid (Addgene 43813) or empty vector control (Addgene 29496).

Techniques: Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Clinical Proteomics, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Wnt5a-mediated Adipo-Cardiac Interorgan Communication in HFpEF

doi: 10.1101/2025.10.29.685456

Figure Lengend Snippet: (A) Schematic outline of study. (B) Histologic and wheat germ agglutinin-based hypertrophy, (C) plasma Wnt5a levels, (D) left ventricular systolic pressure, (E) left ventricular end-diastolic pressure, (F) diastolic relaxation of the left ventricle, (G) right ventricular systolic pressure, (H) right ventricular end-diastolic pressure, (I) diastolic relaxation of the right ventricle, (J) left atrial end-systolic volume by echocardiography, and (K) E/e’ ratio by echocardiography in C57/BL6J mice treated with 8 weeks of L-NAME and high fat diet, followed by LGK974 or carrier control intraperitoneal injections.

Article Snippet: For studies investigating the effects of Wnt5a treatment, cells were transfected with either pcDNA3.2-Wnt5a plasmid (Addgene 43813) or empty vector control (Addgene 29496).

Techniques: Clinical Proteomics, Control

Journal: eLife

Article Title: CRISPR-edited DPSCs constitutively expressing BDNF enhance dentin regeneration in injured teeth

doi: 10.7554/eLife.105153

Figure Lengend Snippet: ( A ) A schematic representation of the transplantation of DPSC into the first molar tooth after drilling. ( B ) The confirmation of BDNF CRISPR activation plasmid enhanced the expression of pro-BDNF. ( C ) Bar graph showing the integrated intensity of CRISPR-engineered BDNF-activated DPSCs against β-actin compared to control. ( D ) H&E staining of sham control and injured tooth in mouse (n = 6 each group) transplanted with CRISPR-engineered BDNF-overexpressing DPSCs. Scale bar: 100 μm. Figure 4—source data 1. Original files for western blot images displayed in . Figure 4—source data 2. Original files for western blot images displayed in with labeling.

Article Snippet: BDNF CRISPR activation plasmid (h) (Cat# sc-400029-ACT) and Reagent System were purchased from Santa Cruz Biotechnology (Dallas).

Techniques: Transplantation Assay, CRISPR, Activation Assay, Plasmid Preparation, Expressing, Control, Staining, Western Blot, Labeling

Journal: eLife

Article Title: CRISPR-edited DPSCs constitutively expressing BDNF enhance dentin regeneration in injured teeth

doi: 10.7554/eLife.105153

Figure Lengend Snippet: ( A ) Immunohistochemistry was performed to assess GFP-tagged transplanted cells. The white arrow indicates the pulp lining. Scale bar: 100 μm ( B ) Micro-CT image in the sham control of the pulp-capping mouse model. The white arrow indicates the drilling operation, and the dotted line specifies the injured area of dentin. The white box is a magnified image of the injured area. ( C ) The transplantation of CRISPR-engineered BDNF-overexpressing DPSCs in the pulp-capping mouse model. ( D ) Analyzed density of dentin compared with sham control vs. transplantation of CRISPR-engineered BDNF-overexpressing DPSCs in the pulp-capping mouse model (n=5). p<0.05 vs. control.

Article Snippet: BDNF CRISPR activation plasmid (h) (Cat# sc-400029-ACT) and Reagent System were purchased from Santa Cruz Biotechnology (Dallas).

Techniques: Immunohistochemistry, Micro-CT, Control, Transplantation Assay, CRISPR

Journal: eLife

Article Title: CRISPR-edited DPSCs constitutively expressing BDNF enhance dentin regeneration in injured teeth

doi: 10.7554/eLife.105153

Figure Lengend Snippet: ( A ) Representative image of sham control of the pulp-capping mouse model. ( a–j ) Sham control vs. ( k–t ) representative image of the transplantation of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-engineered BDNF-overexpressing DPSCs in the pulp-capping mouse model. Scale bars: 25 and 50 μm. ( B ) Line graphs showing the co-localization of BDNF and GFP [B (e1 and j1)] and of TrkB and GFP [B (o1 and t1)].

Article Snippet: BDNF CRISPR activation plasmid (h) (Cat# sc-400029-ACT) and Reagent System were purchased from Santa Cruz Biotechnology (Dallas).

Techniques: Control, Transplantation Assay, CRISPR

Journal: eLife

Article Title: CRISPR-edited DPSCs constitutively expressing BDNF enhance dentin regeneration in injured teeth

doi: 10.7554/eLife.105153

Figure Lengend Snippet: Proposed interaction of tumor necrosis factor alpha (TNFα) and brain-derived neurotrophic factor (BDNF)/tropomyosin receptor kinase B (TrkB) downstream signaling to modulate odontoblastic differentiation in dental pulp stem cells (DPSCs).

Article Snippet: BDNF CRISPR activation plasmid (h) (Cat# sc-400029-ACT) and Reagent System were purchased from Santa Cruz Biotechnology (Dallas).

Techniques: Derivative Assay