Fig 2: siRNA knockdown of candidate antigen expression decreases cell survival and increases apoptosis in human breast cancer cell lines.Four pooled siRNA knocked down expression of the targets VPS35 (a, b), ARPC2 (c, d), SERBP1 (e, f), KRT8 (g, h), and Pdia6 (i, j). Decreased expression of these proteins demonstrate decreased survival (a, c, e, g, and i) and increased apoptosis (b, d, f, h, and j) as compared to mock transfected in either triple negative (TNBC, HCC70) or HER2 positive (HER2, SKBR3) human breast cancer cell lines. Survival is measured as % survival compared to mock transfected cells and apoptosis is measured as fold increase compared to mock transfected calls. All studies are in triplicate. Error bars are standard error of mean (SEM) ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.

Fig 3: Vaccination with antigen-specific peptides was immunogenic and IFN-g immune response correlated with smaller tumor volume.Corrected spots per well (CSPW mean peptide wells as compared to no antigen wells using 3.5 × 105 cells/well) was correlated by linear regression with tumor volume (mm3) for animals that received vaccination with pooled epitopes to a specific antigens and animals that received PBS controls. IFN-g immune response was evaluated by IFN-g ELISPOT. a VPS35, b ARPC2, c SERBP1, d KRT8, and e PDIA6. R2 and p value shown in each panel.

Fig 4: Autoantibodies against the antigens are increased in patients with breast atypia as compared to patients without breast atypia.Indirect ELISA of sera from women without breast atypia (n = 43) was compared to sera from women undergoing breast surgery for mammographic abnormalities (benign pathology n = 12, hyperplasia n = 12, fibroadenoma n = 36, or DCIS n = 59), and women with known breast cancer (n = 37). Y axis is ng/mL IgG antibody concentration by ELISA for each antigen and X axis is the breast pathology a VPS35, b ARPC2, c SERBP1, d KRT8, and e PDIA6 Error bars are standard error of mean (SEM) ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.