Supplementary Figure 1 from RANKL Expression, Function, and Therapeutic Targeting in Multiple Myeloma and Chronic Lymphocytic Leukemia
Year: 2023
Type: supplementary-materials
Abstract: PDF file - 184K, (A) The RANKL antibodies MIH23 and MIH24 specifically bind to RANKL. Binding of MIH23 and MIH24 to surface expressed RANKL was determined by FACS using RANKL transfectants (L-RANKL) and control cells (L cells). Shaded peaks, specific mAb (MIH23 and MIH24); open peaks, isotype controls (mouse IgM and mouse IgG2b, respectively). (B) MIH23 and MIH24 block RANK-RANKL interaction. RANKL transfectants and the parental controls were stained with RANK-Ig (shaded peaks) or human IgG1 (open peaks) followed by anti-human-PE and investigated by FACS. Dotted line: staining after preincubation with the indicated RANKL mAb; dashed line: after preincubation with mouse IgM and mouse IgG2b as respective isotype controls. (C) Sensitivity and specificity of the sandwich ELISA for detection of sRANKL. The indicated serial dilutions of rRANKL in culture medium were analyzed as described in the methods section. X represents 100ng/ml rGITRL as control. The dotted line represents the chosen detection limit of 0.05ng/ml. Means of triplicates with standard deviations are shown. (D, E) MM cell lines do not display RANKL protein on the surface despite expression of mRANKL mRNA. The indicated MM cell lines were analyzed for RANKL surface and mRNA expression by FACS (D) and RT-PCR (E) as described in the methods section. Results obtained with RANKL transfectants and parental cells are shown as controls.
Language: English
Subfield: Hematology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4361955577