mouse recombinant bmp2  (R&D Systems)

Journal: Cell Reports Methods

Article Title: An orthogonal CRISPR/Cpf1 platform for precise spatiotemporal gene regulation and osteoporotic fracture repair

doi: 10.1016/j.crmeth.2025.101299

Figure Lengend Snippet: Multiplex orthogonal gene regulation and programmable editing by OREC with therapeutic application in osteoblasts (A) Experimental illustration of orthogonal activation and repression of ASCL1 and RAB7A gene expression in HEK293T cells using distinct chemical and light inductions. (B) RT-qPCR time course analysis demonstrating orthogonal and reversible control of ASCL1 activation and RAB7A repression in response to alternating Dox and light stimuli. Data represent mean ± SEM ( n = 3). vs. RAB7A baseline (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001); vs. RAB7A 36 h ( $ p < 0.05, $$$ p < 0.001); vs. ASCL1 baseline ( && p < 0.01, &&&& p < 0.0001); vs. ASCL1 48 h ( ## p < 0.01, #### p < 0.0001). (C) Schematic diagram illustrating crRNA guide-length-dependent transcriptional repression and HDR-based gene editing using OREC c system. (D) Transcriptional repression of RAB7A and FZD1 genes following 72 h doxycycline treatment using 15 bp crRNA guides. Data represent mean ± SEM ( n = 3) (∗∗∗∗ p < 0.0001). (E) Luciferase activity analysis following HDR-mediated luciferase knockin using 20 bp crRNA guides. Data represent mean ± SEM ( n = 3) (∗∗∗∗ p < 0.0001). (F) Flow cytometry analysis showing percentage of GFP-positive cells following HDR-mediated GFP knockin. Data represent mean ± SEM ( n = 3) (∗∗∗∗ p < 0.0001). (G) Schematic representation illustrating simultaneous orthogonal activation of Bmp2 and repression of Dkk1 expression in MC3T3-E1 cells. (H) RT-qPCR analysis demonstrating significantly enhanced Bmp2 activation and Dkk1 repression under simultaneous orthogonal induction compared with single-gene regulatory conditions. Data are presented as mean ± SEM ( n = 3). ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. (I) Representative images of alizarin red S (ARS) and alkaline phosphatase (ALP) staining showing significantly enhanced osteogenic differentiation following orthogonal OREC induction compared to single-gene controls. Color code for experimental conditions: white: empty vector control; gray: OREC o construct only; crosshatched: OREC c construct only; red: OREC o + Light (optogenetic activation); green: OREC c + Dox (chemogenetic activation) (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (J) RT-qPCR analysis demonstrating synergistic upregulation of osteogenic marker genes ( Alp , Bglap , and Sp7 ) upon simultaneous Bmp2 activation and Dkk1 repression. Data represent mean ± SEM ( n = 3) (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (K) The CCK-8 assay quantifies viable cells via dehydrogenase-mediated formazan formation measured at 450 nm, showing that OREC treatments do not affect cell viability in MC3T3-E1 cells. Data shown as mean ± SEM ( n = 3). See also and .

Article Snippet: Recombinant BMP2 protein , MedChemExpress , Cat# HY-P7006.

Techniques: Multiplex Assay, Activation Assay, Gene Expression, Quantitative RT-PCR, Control, Luciferase, Activity Assay, Knock-In, Flow Cytometry, Expressing, Staining, Plasmid Preparation, Construct, Marker, CCK-8 Assay

Journal: Cell Reports Methods

Article Title: An orthogonal CRISPR/Cpf1 platform for precise spatiotemporal gene regulation and osteoporotic fracture repair

doi: 10.1016/j.crmeth.2025.101299

Figure Lengend Snippet: AAV-delivered OREC system design and in vivo orthogonal gene regulation in fracture healing (A) OREC o system comprises two vectors encoding dAsCpf1-Activer (upper) and TetR-CIBN-P2A-CRY2-VP16 3 with Bmp2 -targeting crRNA (lower). (B) OREC c system consists of vectors encoding dLbCpf1-Repr (upper) and another vector expressing rtTA-Advanced with Dkk1 -targeting crRNA (lower). See also . All constructs designed within AAV packaging constraints. (C) Experimental setup showing AAV intratibial injection procedure with custom LED patch device and power supply system for localized light delivery. (D) Experimental timeline: AAV intratibial injection 14 days before fracture (day 14), tibial fracture induction (day 0), and blue light treatment (470 nm, twice daily for 30 min) beginning one day post-fracture (day 1) through endpoint (day 42). (E and F) AAV2/9 vector constructs for bioluminescence analysis of OREC activity in vivo . (G) Experimental timeline showing AAV intratibial injection (day 0) followed by 14-day treatment with light stimulation (470 nm, 30 min twice daily) or oral doxycycline (30 mg/kg daily) prior to bioluminescence imaging. (H) Representative bioluminescence images demonstrating inducible luciferase expression at tibial sites by OREC with no detectable signal in distant tissues. (I) RT-qPCR analyses of Bmp2 and Dkk1 expression levels in orthogonally treated fracture sites at days 7 and 21 post-fracture. Data represent mean ± SEM ( n = 6 mice/group) (∗∗ p < 0.01, ∗∗∗∗ p < 0.0001). (J) Analysis of Bmp2 and Dkk1 expression relative to Gapdh using the 2 −ΔCt method in fracture callus tissue 7 days after light or doxycycline induction in OREC-transduced mice. Data represent mean ± SEM ( n = 6 mice per group). (K and L) RT-qPCR analysis of Bmp2 (K) and Dkk1 (L) expression in fracture callus tissue from mice exposed to inducers without OREC constructs. Data represent mean ± SEM ( n = 3 mice per group). See also and .

Article Snippet: Recombinant BMP2 protein , MedChemExpress , Cat# HY-P7006.

Techniques: In Vivo, Plasmid Preparation, Expressing, Construct, Injection, Activity Assay, Imaging, Luciferase, Quantitative RT-PCR

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: Adam12 + fibroblasts predominantly derive from resident fibroblasts in unwounded skin. A, B Immunofluorescence staining analysis for BMP2 in C57BL/6J mice on unwound and wounded back skin at 0dpw, 3dpw, 7dpw, 10dpw and 14dpw. Error bars represent SD (n=5 wounds). Scale bar = 100μm. C Schematic depicting wounding, vehicle injection and 20μg BMP2 inhibitor LDN-193189 2HCl injection of Adam12-tdTomato mice. D, E Flow cytometry analysis of tdTomato on vehicle injection or LDN-193189 2HCl injection wounds of Adam12-tdTomato mice. Error bars represent SD (n=5 wounds). F Schematic depicting tamoxifen induction and wounding of Marker CreERT2 ; R26 LSL-tdTomato mice for lineage tracing. G, H Immunofluorescence staining analysis for tdTomato and Adam12 on wounds of Pdgfra CreERT2 ; R26 LSL-tdTomato , Sm22 CreERT2 ; R26 LSL-tdTomato and Krt5 CreERT2 ; R26 LSL-tdTomato mice at 10dpw. Error bars represent SD (n=8 wounds for Pdgfra CreERT2 ; R26 LSL-tdTomato mice. n=4 wounds for Sm22 CreERT2 ; R26 LSL-tdTomato and Krt5 CreERT2 ; R26 LSL-tdTomato mice). Scale bar = 100μm. Two-tailed Student's paired t-test was used to determine statistical significance in E . One-way ANOVA test was used to determine statistical significance in B and H .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Immunofluorescence, Staining, Injection, Flow Cytometry, Marker, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: BMP2 signaling is required for the lineage commitment of resident dermal fibroblasts into pro-fibrogenic Adam12 + fibroblasts. A Schematic depicting wounding and tamoxifen induction of Pdgfra CreERT2 ;Bmpr2 fl/fl mice for fibroblasts-specific Bmpr2 deletion. B, C Immunofluorescence staining analysis for Adam12 on wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). Scale bar = 100μm. D, E Representative photographic images of wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 0dpw and 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). F, G Masson staining on wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). Scale bar = 100μm. H, I Immunofluorescence staining analysis for periostin on wound tissues of Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ , and Pdgfra CreERT2 ;Bmpr2 fl/fl mice at 14dpw. Error bars represent SD (n=6 wounds for Pdgfra CreERT2 ;Bmpr2 +/+ , Pdgfra CreERT2 ;Bmpr2 fl/+ and n=8 wounds for Pdgfra CreERT2 ;Bmpr2 fl/fl ). Scale bar = 100μm. One-way ANOVA test was used to determine statistical significance in C, E, G and I .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Immunofluorescence, Staining

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: Augmentation of BMP2 signaling drives expansion of Adam12 + fibroblasts and exacerbates skin scarring. A-F, Immunofluorescence staining analysis for BMP2, ADAM12 and Periostin in normal scar, hypertrophic scar (HTS) and keloid. Error bars represent SD (n=8). Scale bar = 100μm. G The correlation analyses between the expression of BMP and ADAM12. The range between the two dashed lines represents the 95% confidence interval. H The correlation analyses between the expression of ADAM12 and periostin. The range between the two dashed lines represents the 95% confidence interval. I Schematic depicting wounding, 2mg tamoxifen induction, 200ng DT injection and 25ng BMP2 injection of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. J, K Flow cytometry analysis of tdTomato on vehicle+PBS injection, BMP2+PBS injection and BMP2+DT injection wounds of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. Error bars represent SD (n=6 wounds). L, M Representative photographic images of wound tissues of vehicle+PBS group, BMP2+PBS group and BMP2+DT group Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice at 0dpw and 14dpw. Error bars represent SD (n=6 wounds). N, O Masson staining on vehicle+PBS injection, BMP2+PBS injection and BMP2+DT injection wounds of Adam12 CreERT2 ; R26 LSL-tdTomato-2A-DTR mice. Error bars represent SD (n=6 wounds). Scale bar = 100μm. N Schematic illustration showing that abnormal high-expressed BMP2 in healed wound may lead to pathologic scars. One-way ANOVA test was used to determine statistical significance in B, D, F, K, M and O . Pearson correlation was used to measures the strength and direction of a linear relationship between two variables in G and H .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Immunofluorescence, Staining, Expressing, Injection, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: BMP2-induced Adam12 + Fibroblasts Dictate Wound-associated Skin Scarring and Fibrosis

doi: 10.7150/ijbs.123725

Figure Lengend Snippet: BMP2 represents a promising therapeutic target for pathological scars. A Schematic depicting wounding, vehicle injection and 20μg BMP2 inhibitor LDN-193189 2HCl injection of tension induced hypertrophic scar mice model. B Representative photographic images of scar tissues of vehicle injection or LDN-193189 2HCl injection group in tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 5mm. C, D Immunofluorescence staining analysis for Adam12 on vehicle injection or LDN-193189 2HCl injection wounds of tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 100μm. E, F Masson staining on vehicle injection and LDN-193189 2HCl injection wounds of tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 100μm. G-L, Immunofluorescence staining analysis for Collagen I, Collagen III and periostin on vehicle injection and LDN-193189 2HCl injection wounds of tension induced hypertrophic scar mice model. Error bars represent SD (n=5 wounds). Scale bar = 100μm. Two-tailed Student's unpaired t-test was used to determine statistical significance in D, F, H, J and L .

Article Snippet: The detailed protocols of 20mg/mL tamoxifen(T5648, Sigma), 4μg/mL diphtheria toxin (D0564, Sigma), 20μg/mL periostin (2955-F2-050, R&D Systems), 400μg/mL LDN-193189 2HCl (7507, Selleck) and 0.5μg/mL BMP2 (355-BM-010, R&D Systems) injection were described in the main text or figure legends.

Techniques: Injection, Immunofluorescence, Staining, Two Tailed Test