PMC

A. Cell viability of C4-2B Neo control cells and KD_β2-M cells in response to: taxotere (0.3 µM), cisplatin (10 µM) and PS341 (1 µM). (***p<0.001, Student’s t test) B. Cell viability of DU154 prostate cancer cells in response to combination of anti-β2-M Ab (0.5 µg/ml) and cisplatin (100 µM)/doxorubicin (100 µM). (***p<0.001, Student’s t test). C. Cell viability of PC-3 prostate cancer cells in response to combination of anti-β2-M Ab (1 µg/ml) and cisplatin (100 µM). (***p<0.001, Student’s t test).

A. Radiation sensitivity of ARCaP_E and ARCaP_M prostate cancer cells by clongenic assay. (** p<0.01, *** p<0.001, Student’s t test). B. ARCaP_M cells are sensitized to radiation in the presence of anti-β2-M Ab using clongenic assay. (*p<0.03, *** p<0.008, ANOVA). C. Effect of anti-β2-M Ab (0.8 mg/kg) and radiation (15 Gy) on tumor growth in subcutaneous ARCaP_M xenograft nude mice model. (** p<0.01, Student’s t test).

Tumor incidence in mice tibias were analyzed from H&E images and x-ray scans. The tumorigenecity of control group was 94% (n = 18 tibias) and the tumorigenecity of anti-β2-M Ab+irradiation (IR) treated group was 67% (n = 18 tibias). A. Tumor progression in control and combination treatment (anti-β2-M Ab and irradiation) analyzed by PSA measurements (ng/ml). (***p<0.006, Student’s t test). B. Immunohistochemical staining of tibias in control and combination treatment group stained for H&E, β2-M protein, iron staining, p-CREB and p-histone H3 (10X).

A. Iron staining in control and anti-β2-M Ab (5 µg/ml) treated cells using iron staining kit in ARCaP_M prostate cancer cell lines. B. Mitochondrial superoxide levels in response to anti-β2-M Ab treatment in a time and dose dependent manner in i. ARCaP_M, ii. ARCaP_E prostate cancer cell lines (***p<0.001, Student’s t test) and iii. p69 immortalized prostate epithelial cells using MitoSOX dye. C. Mitochondrial superoxide in ARCaP_M, KD_HFE1 and KD_HFE3 using MitoSOX dye (*p<0.05, Student’s t test). D. Expression of stress response proteins and DNA repair enzymes in C4-2B Neo control and β2-M knockdown cell lines. i.β2-M protein expression, ii. HSP27 and HSP70 protein expression and iii. NUDT1 and MPG protein expression. NUDT1 and MPG protein expression in response to anti- β2-M Ab treatment in LNCaP and C4-2 prostate cancer cells.

A. Cell viability of TRAMP C1 and TRAMP C2 prostate cancer cells in response to anti-β2-M Ab. (***p<0.001, Student’s t test). B. Merged near infra-red and X-ray image of abdomen of TRAMP mice treated with control IgG and anti-β2-M Ab (n = 4). Representative parental mice used as additional control (C57BL/6 mice). The tumorigenecity of control IgG antibody group was 100% (n = 4) and the tumorigenecity of anti-β2-M Ab treated group was 25% (n = 4). C. H&E images of prostates of control IgG mice and anti-β2-M Ab treated mice (10X). D. Immune cell (T and B cells) numbers of wild type mice, control IgG mice and anti-β2-M Ab treated mice measured by flow cytometry. E. Body weights of TRAMP mice treated with IgG or anti-β2-M Ab.