human recombinant bmp2  (PeproTech)

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: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Over-expressed Phospholamban in Cancer-associated Fibroblasts Is a Driver for CMS4 Subtype Colorectal Cancer Formation

doi: 10.1016/j.jcmgh.2025.101524

Figure Lengend Snippet: ( A ) qPCR analysis of angiogenesis marker genes mRNA levels in HUVEC cells qPCR analysis of angiogenesis marker genes mRNA levels in HUVEC cells incubated with CM from NFs transfected with empty vector, PLN overexpression (OE) and with/without anti-GREM1 neutralizing antibody (Ginisortamab, 10 μg/mL) for 48 hours. ( B ) qPCR assay for EMT-related genes of HCT116 and HT29 cells after BMP2, BMP4, BMP7 overexpressed or 20 ng/mL rBMP2, rBMP4, rBMP7 treatments with CAF CM incubation. ( C–D ) Representative photos and quantification of Transwell assay for HCT116 and HT29 cells after BMP2 overexpressed or 20 ng/mL rBMP2 treatment with CAF CM incubation. ( E–F ) Representative photos and quantification of wound healing assay for HCT116 and HT29 cells after BMP2 overexpressed or 20 ng/mL rBMP2 treatment with CAF CM incubation. ( G–H ) representative images of Transwell assay and quantification for cell invasion rates in empty vector or BMP2 OE transfected CT26, or wildtype CT26 treated with rBMP2 (20 ng/mL), all under incubation with mouse CAFs CM for 48 hours. ( I–K ) qPCR assay for angiogenesis-related genes of HUVEC cells after BMP2, BMP4, BMP7 overexpressed or 20 ng/mL rBMP2, rBMP4, rBMP7 treatments with CAF CM incubation. ( L–O ) qPCR assay for angiogenesis-related genes, as well as representative photos and quantification of tube formation assay in HUVEC cells after VEGFR2 knockdown with CAF CM incubation. ( P–R ) Representative images of tube formation assay ( P ), quantification of branching points ( Q ), and tube length ( R ) in C166 cells transfected with shCtrl, shVEGFR2 1#, or shVEGFR2 2# and incubated with mouse CAFs CM for 48 hours. ( S ) relative mRNA levels of PLN in human NFs after incubation with CM from NCM460 (normal colon epithelial cells)、primary CRC cancer cells (prCRC), and metastasis CRC cancer cells (mCRC) for 48 hours, as quantified by qPCR. ( T ) Representative Western blotting bands and quantification of PLN protein levels in NFs treated with CM from NCM460, prCRC, and mCRC. All experiments have been repeated at least 3 times independently. n.s., no significant differences; ∗∗ P < .01.

Article Snippet: Recombinant human BMP2 , Proteintech , HZ-1128.

Techniques: Marker, Incubation, Transfection, Plasmid Preparation, Over Expression, Transwell Assay, Wound Healing Assay, Tube Formation Assay, Knockdown, Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Over-expressed Phospholamban in Cancer-associated Fibroblasts Is a Driver for CMS4 Subtype Colorectal Cancer Formation

doi: 10.1016/j.jcmgh.2025.101524

Figure Lengend Snippet: ( A–I ) qPCR analysis of efficiency BMP2, BMP4, and BMP7 overexpression in HCT116, HT29 and HUVEC cells. ( J ) qPCR analysis of shRNA interference efficiency targeting VEGFR2 in HUVEC cells. ( K–N ) qPCR analysis of shRNA interference efficiency targeting ITGB1, MET, EFBN1, and CSF1R in human NFs. ( O–S ) qPCR analysis of shRNA interference efficiency targeting COL3A1, VTN, HGF, EPHA6, and CSF1 in mCRC cells. All experiments have been repeated at least 3 times independently. n.s., no significant differences; ∗∗ P < .01.

Article Snippet: Recombinant human BMP2 , Proteintech , HZ-1128.

Techniques: Over Expression, shRNA

Journal: PLOS ONE

Article Title: IRX-related homeobox gene MKX is a novel oncogene in acute myeloid leukemia

doi: 10.1371/journal.pone.0315196

Figure Lengend Snippet: (A) Genomic copy number analysis of OCI-AML3 showing the complete chromosome 10 (above), and the chromosomal region 10p12.1-p12.3 highlighting MLLT10 and MKX (below). (B) Data obtained from the UCSC genome browser indicating a CpG-island at MKX (left). RQ-PCR analysis of MKX and NANOG in AML cell line HL-60 treated with DNA-methyltransferase inhibitor AZA (right). The expression level of HL-60 control was set as 1 while the remaining samples are related to that value. (C) RQ-PCR analysis of IRX3 and MKX in OCI-AML3 treated with BMP2 (left) and dorsomorphin (middle). RQ-PCR analysis of SMAD4 , IRX3 and MKX in OCI-AML3 treated for siRNA-mediated knockdown (right). (D) RQ-PCR analysis of IRX3 and MKX (left), and IRX5 and MKX (right) in OCI-AML3 treated for siRNA-mediated knockdown. (E) RQ-PCR analysis of JUNB and MKX in OCI-AML3 treated for siRNA-mediated knockdown. The expression level of OCI-AML3 siCTR was set as 1 while the remaining samples are respectively related to that value. Statistical significance was assessed by Student´s T-Test (two-tailed) and the resultant p-values indicated by asterisks (* p<0.05, ** p<0.01, *** p<0.001, n.s. not significant).

Article Snippet: Additional cell treatments were performed using 20 ng/ml recombinant BMP2 or TNFSF11 (R & D Systems, Wiesbaden, Germany), 100 μM azacytidine (AZA), 10 μM dorsomorphin, 14 μM NFkB-inhibitor, and 100 μM etoposide (Sigma-Aldrich, Taufkirchen, Germany) for 20 h. For functional testing, treated cells were analyzed using the IncuCyte S3 Live-Cell Imaging Analysis System and the Cell-by-Cell analysis software (Sartorius, Göttingen, Germany).

Techniques: Expressing, Control, Knockdown, Two Tailed Test