Fig. 4: αHER2-eStcE is effective in mixed cell assays and breast cancer mouse models.
From: Design of a mucin-selective protease for targeted degradation of cancer-associated mucins
a, Setup for mixed cell suspension survival assays under anchorage-free conditions as in Fig. 1b. b, Viability of anchorage-free mixed MCF10AMUC1, ±HER2 cells ± 1 nM StcE or αHER2-eStcE over 72 h as determined by flow cytometry (n = 3 biologically independent replicates). c, Setup for mixed cell NK cell killing assay as in Fig. 1d. d, Normalized NK cell killing of mixed K562±HER2 cells treated with conjugate at a 2:1 effector:target ratio as determined by flow cytometry (n = 3 biologically independent replicates). Values are reported as fold change relative to the average of three PBS-treated control replicates (dotted line). e, Treatment regimen for BALB/c mice injected i.v. via tail vein with 4T07MUC1, HER2 cells. αHER2-eStcE was injected i.v. every other day at 10 mg per kg starting on day 2 (n = 7 animals per group). f, Plot depicting lung masses of animals described in e. g, Percent area of lung metastases quantified by H&E tissue staining of animals described in e. Points represent individual scans of lung slices (n = 2 per animal). For images, see Extended Data Fig. 9f,g. h, Treatment regimen for BALB/c mice injected with EMT6HER2 orthotopically into the mammary fat pad. αHER2-eStcE or αHER2 was injected four times i.p. every other day starting on day 8 (n = 9 animals per group (αHER2), 10 animals per group (αHER2-eStcE) or 12 animals per group (untreated)). Mice were killed once tumor size reached approximately 1,500 mm3 or when mice developed ulcerated tumors. i, Average growth curves of EMT6HER2 tumors for animals described in h. j, Survival curves for animals described in h. Data are shown as mean ± s.e.m. (b, f, g and i) or mean ± s.d. (d). P values were determined by Tukey-corrected two-way ANOVA (d and i), two-tailed Mann–Whitney test (f and g) and Mantel–Cox test (j); *P < 0.05; **P < 0.005; ***P < 0.0005.
