Anti-Human TNFRSF10A Recombinant Antibody (Mapatumumab) (CAT#: TAB-H48)

Figure 1 Mapatumumab and etoposide synergize to induce apoptosis through the mediation of NF-jB.

(A) Inhibition of NF-jB disrupts synergy between Mapatumumab and etoposide. HT29 cells were untreated, treated with 100 lM etoposide, 10 lM BAY 11-7082, or 100 ug/ml of Mapatumumab alone or in combination. 48 h after treatment cells were collected and cell death was assessed by acridine orange staining. Error bars represent the standard deviation of three independent experiments. Statistical significance was calculated using a Student's t test. (B) Combination treatment between Mapatumumab and Etoposide increases DNA fragmentation; this is disrupted by NFjB blockage. Cells were treated with 100 uM etoposide, 10 lM BAY 11-7082, 100 ug/ml of Mapatumumab, or 20 uM of z-VAD-FMK alone or in combination. 48 h after transfection cells were collected and stained with propidium iodide staining buffer.

Mendoza, F. J., Ishdorj, G., Hu, X., & Gibson, S. B. (2008). Death receptor-4 (DR4) expression is regulated by transcription factor NF-κB in response to etoposide treatment. Apoptosis, 13(6), 756.

Figure 2 Cleaved caspase-8 was associated with DR4 or DR5 in relation to mapatumumab or lexatumumab sensitivity.

Immunoprecipitation (IP) assays of mapatumumab (Mapa) or lexatumumab (Lexa) were conducted in LP1, MDN, and NCI-H929 cells. Cells (20 × 106) were incubated with or without (control) 10 μg/mL of mapatumumab (45 minutes) or lexatumumab (90 minutes) at 37°C in RPMI-1640 containing 5% FCS. For NCI-H929, cells were pretreated for 2 hours with 2.5μmol/L nutlin-3a (N) before addition of lexatumumab. The lysates were immunoprecipitated with a mix of proteins A and G (2 mg of lysate proteins were immunoprecipitated in each assay). For untreated cells, IP was conducted after addition of mapatumumab or lexatumumab to the lysates. The immunoprecipitates were analyzed for the presence of the indicated proteins by immunoblot (IB) analysis. Input, lysates of untreated or treated cells; Ct Ip< IP of control lysates (untreated cells); Ip< IP of lysates obtained from cells treated with mapatumumab or lexatumumab. *, heavy chain of mapatumumab or lexatumumab.

Surget, S., Chiron, D., Gomez-Bougie, P., Descamps, G., Ménoret, E., Bataille, R., ... & Pellat-Deceunynck, C. (2012). Cell death via DR5, but not DR4, is regulated by p53 in myeloma cells. Cancer research, canres-0487.