Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology
Article Title: Inhibition of the stem cell factor 248 isoform attenuates the development of pulmonary remodeling disease
doi: 10.1152/ajplung.00114.2019
Figure Lengend Snippet: Characterization of anti-SCF248 mAb. A: using ATCC cell lines CRL-2452, CRL-2453, and CRL-2454 that express no SCF, SCF220, or SCF248, respectively, the monoclonal antibody was used for flow cytometry binding assays to demonstrate specificity. B: monoclonal antibody generated to a peptide from exon 6 of SCF248 was subjected to Biacore surface plasmon resonance (SPR) analysis to the specific peptide. The data generated dose response curves that were assessed to have a fast on rate and a slow off rate with a 4.5 nM KD binding. C: image flow cytometry photos of ATCC SCF248-expressing cells incubated for 5 or 60 min with anti-SCF248 or control IgG1 mAb coupled with Phrodo-red pH-sensitive fluorescent dye. D: mean fluorescent intensity (MFI) of ATCC SCF248-expressing cells incubated with control or anti-SCF248 mAb coupled with phrodo-red dye over time to demonstrate internalization. E: IPF patient lung fibroblast cultures incubated for 15 min with control or Anti-SCF248 mAb coupled with phrodo-red dye showing internalization only in the anti-SCF48 mAb-incubated cells. F: naïve Balb/c/J mice were injected intravenously with either control IgG (250 mg/kg), polyclonal anti-SCF (250 mg/kg), or anti-SCF248 monoclonal ab (100 mg/kg) on day 0, 2, 4, and 6. The peripheral blood was assessed for reticulocyte numbers on day 8 as an indication of reduced erythropoiesis as a % of total cells. Data represent the mean ± SE from eight mice/group. ATCC, American Type Culture Collection; IPF, idiopathic pulmonary fibrosis; SCF, stem cell factor.
Article Snippet: A : using ATCC cell lines CRL-2452, CRL-2453, and CRL-2454 that express no SCF, SCF220, or SCF248, respectively, the monoclonal antibody was used for flow cytometry binding assays to demonstrate specificity.
Techniques: Flow Cytometry, Binding Assay, Generated, SPR Assay, Expressing, Incubation, Injection