Flow cytometry Following the dissection of soft tissue, the knee joints and blood were harvested. For the analysis of subchondral endothelial cells and MSCs, 10 subchondral bone specimens from 5 knee joints (including tibial and femoral subchondral bone in each knee joint) were used for cell isola...
Immunoblotting, flow cytometry, cell viability test, and confocal microscopy were conducted to assess the internalization and cytotoxic effect of ADC targeting mPDGFRβ in an in vitro setting. Immunofluorescence staining of whole-mount retinas and retinal pigment epithelium-choroid-scleral complexes, elect...
For many, if not most, proteins of interest, such high quality antibodies are not available. The ability to convert any biological process or cell state into a completely generic gene expression signature that can be monitored in high throughput and at low cost is therefore attractive. The ...
Specifically, we examined the influence of inflammation on platelet-derived growth factor receptor-beta (PDGFRβ) expression and signalling, as well as pericyte proliferation and migration by qRT-PCR, immunocytochemistry, flow cytometry, and western blotting. Results Chronic IFNγ treatment had marked ...
staining. List of antibodies for cell surface markers (name of the clone) for human flow cytometry: CD3 (SK7 or UCHT1), CD20 (2H7), CD4 (RPA-T4), CD8 (RPA-T8), CD14 (61D3), CCR6 (11A9), CD27 (M-T271) and CD43 (1G10) were purchased from eBioscience and BD Bioscience. ...
1 min at RT. Single-cell suspensions were incubated for 15 min in 1% FBS-PBS and stained with the following fluorescence-conjugated antibodies (Additional file1: Table S1). Samples were acquired on a LSR IV Fortessa Flow Cytometer (BD Biosciences). Data were analyzed on FlowJo V10 (...
Flow-cytometry Single cell suspension of primary SCs and MFs (1 × 106 cells/500 μl of 1% FCS containing DMEM) were stained by incubating with primary antibodies, directly conjugated with fluorochrome at 4 °C for 30 min followed by washing with PBS-BSA solution. For biotinylated and ...
A, B, and D–I Cell or tissue lysates were subjected to immunoblotting with the indicated antibodies. Error bars indicate mean ± SD of triplicate samples. ****P < 0.0001. Full size image Due to the activation of AKT1 leading to both an increase in CREB activity and PDGFRα ...
After pretreatment with baicalin, VSMCs were stimu- lated with PDGF-BB for 30 min, and the phosphoryla- tion status of ERK, c-Jun NH2-terminal kinase (JNK), p38 MAPK, and Akt was measured by western blot analysis using antibodies that identify the active (phos- phorylated) forms of ...
Then, proteins were incubated with primary antibodies for 16 h at 4 ◦C by gentle rocking using different concentrations (usually 1:500 for phosphoantibodies and 1:1000 for the rest of the antibodies), depending on the antibody against PDGFR-α (sc-431), PDGFR-β (sc-432), phospho-...