Journal: Scientific Reports

Article Title: MicroRNA-378a-3p is overexpressed in psoriasis and modulates cell cycle arrest in keratinocytes via targeting BMP2 gene

doi: 10.1038/s41598-021-93616-8

Figure Lengend Snippet: Validation of candidate gene targets of miR-378a. ( a ) The expression levels of BMP2 and INHBA by qPCR using mRNA from NHEK cells transfected with miR-378a mimic or inhibitor for 24 h and 48 h. ( b ) Comparison of the expression levels of the selected candidate genes from high-throughput RNA-sequencing (light colors) versus real-time qPCR (dark colors) from NHEK cells over-expressing miR-378a mimic (red) or inhibitor (blue) for 24 h and 48 h. Error bars indicate mean ± SEM from three replicates. The experiment was performed three times independently. *Indicates P < 0.05, **indicates P < 0.01, and ***indicates P < 0.001.

Article Snippet: The membranes were cut according to molecular weight of the proteins and initially blocked with blocking buffer (5% non-fat dry milk in PBST) for 1 h. Later, it was immunoblotted with anti-BMP2 antibody (1:1000 dilution; #AHP2442, Bio-Rad Laboratories, Inc.), anti-involucrin antibody (1:1000 dilution; #sc-398952, Santa Cruz Biotechnology), and anti-GAPDH (1:5000 dilution; Santa Cruz Biotechnology, Dallas, TX, USA; cat#sc-25778) for 1 h at room temperature.

Techniques: Biomarker Discovery, Expressing, Transfection, Comparison, High Throughput Screening Assay, RNA Sequencing

Journal: Scientific Reports

Article Title: MicroRNA-378a-3p is overexpressed in psoriasis and modulates cell cycle arrest in keratinocytes via targeting BMP2 gene

doi: 10.1038/s41598-021-93616-8

Figure Lengend Snippet: BMP2 as a novel target gene of miR-378a in keratinocytes. ( a ) Hypothetical binding structure of miR- 378a at 3′UTR of BMP2 (left) and INHBA (right) with predictive binding energy and probability value for actual binding. ( b ) Luciferase activity of wild-type and mutant 3′UTR of BMP2 genes in HEK293T cells co-transfected with miR-378a mimic or inhibitor for 48 h. ( c ) Western blot images (top) and quantitative measurement (bottom) of BMP2 protein in NHEK cells with miR-378a overexpression or suppression for 72 h. A representative image of BMP2 protein from four independent experiments was shown (also see Supplementary Fig. ). ( d ) The expression level of BMP2 in psoriatic lesions (n = 4) and healthy individuals (n = 4). *Indicates P < 0.05, **indicates P < 0.01, and ***indicates P < 0.001.

Article Snippet: The membranes were cut according to molecular weight of the proteins and initially blocked with blocking buffer (5% non-fat dry milk in PBST) for 1 h. Later, it was immunoblotted with anti-BMP2 antibody (1:1000 dilution; #AHP2442, Bio-Rad Laboratories, Inc.), anti-involucrin antibody (1:1000 dilution; #sc-398952, Santa Cruz Biotechnology), and anti-GAPDH (1:5000 dilution; Santa Cruz Biotechnology, Dallas, TX, USA; cat#sc-25778) for 1 h at room temperature.

Techniques: Binding Assay, Luciferase, Activity Assay, Mutagenesis, Transfection, Western Blot, Over Expression, Expressing

Journal: International Journal of Nanomedicine

Article Title: A Three-Dimensional Printed Polycaprolactone Scaffold Combined with Co-Axially Electrospun Vancomycin/Ceftazidime/Bone Morphological Protein-2 Sheath-Core Nanofibers for the Repair of Segmental Bone Defects During the Masquelet Procedure

doi: 10.2147/IJN.S238478

Figure Lengend Snippet: SEM image and fiber diameter distribution of ( A ) pure PLGA nanofibers, ( B ) sheath-core PLGA/BMP-2 nanofibers, and ( C ) PLGA/vancomycin/ceftazidime nanofibers.

Article Snippet: Commercial antibodies were adapted for IHC analysis; these included anti-BMP-2 (polyclonal, 1:200; cat. no. A0231; ABclonal, MA, USA); anti-VEGF (polyclonal, 1:200; cat. no. A0280; ABclonal); anti-vWF (vWF Picoband antibody, 1:200; cat. no. PB9062; Boster Biological Technology, Pleasanton CA, USA); anti-TGF-β (polyclonal, 1:200; cat. no. A2561; ABclonal); and anti-IL-6 (polyclonal, 1:200; cat. no. A0286; ABclonal).

Techniques:

Journal: International Journal of Nanomedicine

Article Title: A Three-Dimensional Printed Polycaprolactone Scaffold Combined with Co-Axially Electrospun Vancomycin/Ceftazidime/Bone Morphological Protein-2 Sheath-Core Nanofibers for the Repair of Segmental Bone Defects During the Masquelet Procedure

doi: 10.2147/IJN.S238478

Figure Lengend Snippet: Water contact angles of ( A ) pure PLGA nanofibers (126.24 ± 10.26°), ( B ) sheath-core PLGA/BMP-2 nanofibers (120.00 ± 8.67°), and ( C ) PLGA/vancomycin/ceftazidime nanofibers (69.2.33 ± 5.14°). Substantially improved hydrophilicity of electrospun nanofibers was observed with the addition of water-soluble antibiotics.

Article Snippet: Commercial antibodies were adapted for IHC analysis; these included anti-BMP-2 (polyclonal, 1:200; cat. no. A0231; ABclonal, MA, USA); anti-VEGF (polyclonal, 1:200; cat. no. A0280; ABclonal); anti-vWF (vWF Picoband antibody, 1:200; cat. no. PB9062; Boster Biological Technology, Pleasanton CA, USA); anti-TGF-β (polyclonal, 1:200; cat. no. A2561; ABclonal); and anti-IL-6 (polyclonal, 1:200; cat. no. A0286; ABclonal).

Techniques:

Journal: International Journal of Nanomedicine

Article Title: A Three-Dimensional Printed Polycaprolactone Scaffold Combined with Co-Axially Electrospun Vancomycin/Ceftazidime/Bone Morphological Protein-2 Sheath-Core Nanofibers for the Repair of Segmental Bone Defects During the Masquelet Procedure

doi: 10.2147/IJN.S238478

Figure Lengend Snippet: In vitro drug release profiles. ( A ) Daily and ( B ) accumulated release of antibiotics from the composite nanofibers. ( C ) Daily and ( D ) accumulated release of BMP-2 from the composite nanofibers. In vivo examination of the daily release of ( E ) antibiotics and ( F ) BMP-2 from the composite scaffold in the muscular tissue.

Article Snippet: Commercial antibodies were adapted for IHC analysis; these included anti-BMP-2 (polyclonal, 1:200; cat. no. A0231; ABclonal, MA, USA); anti-VEGF (polyclonal, 1:200; cat. no. A0280; ABclonal); anti-vWF (vWF Picoband antibody, 1:200; cat. no. PB9062; Boster Biological Technology, Pleasanton CA, USA); anti-TGF-β (polyclonal, 1:200; cat. no. A2561; ABclonal); and anti-IL-6 (polyclonal, 1:200; cat. no. A0286; ABclonal).

Techniques: In Vitro, In Vivo

Journal: International Journal of Nanomedicine

Article Title: A Three-Dimensional Printed Polycaprolactone Scaffold Combined with Co-Axially Electrospun Vancomycin/Ceftazidime/Bone Morphological Protein-2 Sheath-Core Nanofibers for the Repair of Segmental Bone Defects During the Masquelet Procedure

doi: 10.2147/IJN.S238478

Figure Lengend Snippet: Analysis of the gross specimen ( A ) revealed a thick induced membrane (IM) formed circumferentially around the applied scaffold. The PLGA nanofibers had been dissolved completely, and only the PCL mesh was preserved. The histological evaluation of the induced membrane was performed (blue arrow) and represents in ( B ). ( B ) Histological evaluation by hematoxylin and eosin staining of the induced membrane. Radiographic examination of fracture healing in ( C ) the PCL group, ( D ) the PCL-PLGA/antibiotic group, and ( E ) the PCL-PLGA/antibiotic/BMP-2 group.

Article Snippet: Commercial antibodies were adapted for IHC analysis; these included anti-BMP-2 (polyclonal, 1:200; cat. no. A0231; ABclonal, MA, USA); anti-VEGF (polyclonal, 1:200; cat. no. A0280; ABclonal); anti-vWF (vWF Picoband antibody, 1:200; cat. no. PB9062; Boster Biological Technology, Pleasanton CA, USA); anti-TGF-β (polyclonal, 1:200; cat. no. A2561; ABclonal); and anti-IL-6 (polyclonal, 1:200; cat. no. A0286; ABclonal).

Techniques: Staining

Journal: International Journal of Nanomedicine

Article Title: A Three-Dimensional Printed Polycaprolactone Scaffold Combined with Co-Axially Electrospun Vancomycin/Ceftazidime/Bone Morphological Protein-2 Sheath-Core Nanofibers for the Repair of Segmental Bone Defects During the Masquelet Procedure

doi: 10.2147/IJN.S238478

Figure Lengend Snippet: Immunohistochemical analysis of the expression of ( A ) BMP-2 (*membrane-lining cells, † spindle cells), ( B ) TGF-β (*membrane-lining cells, † spindle cells), ( C ) vWF, ( D ) VEGF, and ( E ) IL-6.

Article Snippet: Commercial antibodies were adapted for IHC analysis; these included anti-BMP-2 (polyclonal, 1:200; cat. no. A0231; ABclonal, MA, USA); anti-VEGF (polyclonal, 1:200; cat. no. A0280; ABclonal); anti-vWF (vWF Picoband antibody, 1:200; cat. no. PB9062; Boster Biological Technology, Pleasanton CA, USA); anti-TGF-β (polyclonal, 1:200; cat. no. A2561; ABclonal); and anti-IL-6 (polyclonal, 1:200; cat. no. A0286; ABclonal).

Techniques: Immunohistochemical staining, Expressing

Journal: Molecular Medicine Reports

Article Title: Dose-dependent inhibitory effects of zoledronic acid on osteoblast viability and function in vitro

doi: 10.3892/mmr.2015.4627

Figure Lengend Snippet: Effect of ZA on the expression of BMP-2 and phosphorylation of the ERK 1/2 and p38 pathways in MC3T3-E1 cells. The MC3T3-E1 cells were incubated with various concentrations of ZA (0–1 µ M) for 7 days. (A) Levels of secreted BMP-2, measured using an ELISA. (B) mRNA expression levels of BMP-2, determined using reverse transcription-quantitative polymerase chain reaction analysis. (C) Protein levels of BMP-2, p38, p-p38, ERK 1/2 and p-ERK 1/2, detected using western blot analysis and normalized to GAPDH. (D) Quantitative analysis of the blots for BMP-2. (E) Relative expression of p-ERK, normalized to ERK, and relative expression of p-p38 normalized to p38. The results are expressed as the mean ± standard error of the mean (n =3 for each group). * P<0.05, ** P<0.01 and *** P<0.001, compared with the 0 µ M group. BMP-2, bone morphogenetic protein 2; ERK 1/2, extracellular signal-regulated kinase 1/2; p-, phosphorylated; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Primary antibodies against the following targets were used: Monoclonal rabbit anti-p38 antibody (1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA; cat. no. 8690), monoclonal rabbit anti-phosphorylated (p)-p38 antibody (1:1,000; Cell Signaling Technology, Inc.; cat. no. 4511), monoclonal rabbit anti-ERK 1/2 antibody (1:1,000; Cell Signaling Technology, Inc.; cat. no. 4695), monoclonal rabbit anti-p ERK 1/2 antibody (1:1,000; Cell Signaling Technology, Inc.; cat. no. 4370), polyclonal rabbit anti-inactive caspase-3 antibody (1:500; Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. SC-7148), polyclonal rabbit anti-OCN antibody (1:500; Santa Cruz Biotechnology, Inc.; cat. no. SC-30045), polyclonal rabbit anti-active caspase-3 antibody (1:200; Abcam, Cambridge, UK; cat. no. ab2302), monoclonal rabbit anti-ALP antibody (1:20,000; Abcam; cat. no. ab108337), polyclonal rabbit anti-BMP-2 antibody (1:1,000; Abgent Biotech Co., Ltd., Suzhou, China; cat. no. AP13858c), polyclonal rabbit anti-Runx2 antibody (1:400; Wuhan Boster Biological Technology, Ltd.; cat. no. BA3613-2), monoclonal mouse anti-glyceraldehyde phosphate dehydrogenase (GAPDH) antibody (1:400; Wuhan Boster Biological Technology, Ltd.; cat. no. BM1623), monoclonal mouse anti-β actin antibody (1:400; Wuhan Boster Biological Technology, Ltd.; cat. no. BM0627).

Techniques: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot