Necroptosis in Both Tumour and Stromal Compartments Determines Responsiveness to Immunogenic Cell Death-based Immunotherapy

Updates to Raw Data Files (Zenodo Version 2) Fig. 4E-For the cell death assay dataset, values previously reported for individual three-dimensional fields of view (8 total fields) have been consolidated at the well level in accordance with journal requirements. Specifically, two fields per well were averaged to generate one value per well, resulting in four technical replicate values (one per independent well). Fig. 4F-Western blot annotations have been revised to accurately indicate first and re-probed membranes. Fig. 4G, H and I-For these dataset, measurements (4 individual three-dimensional fields derived from 2 wells) have been averaged per well (two fields per well), resulting in two technical replicate values (one per independent well), in accordance with journal requirements. Fig. 5N-In accordance with journal requirements, for immune cell quantification datasets, section-level counts were re-analysed to standardise prior to averaging per tumour. Each value represents one independent tumour. Fig. 5P- In accordance with journal requirements, each value represents one independent tumour. Fig. 6C-For these data, measurements have been averaged per well, resulting in three technical replicate values (one per independent well), in accordance with journal requirements. Fig. 6B_BP487 O-Western blot annotations have been revised to accurately indicate first and re-probed membranes. Specifically, GAPDH is a first blot and not a re-blot. Image data remain unchanged. Fig. 6B_BP903 O-Western blot annotations have been revised to accurately indicate first and re-probed membranes. Specifically: a-IRF-3 blot is a re-blot not a first blot and aP-STAT3 blot is first blot, not a re-blot. Image data remain unchanged. Fig. 6M-In accordance with journal requirements, each value represents one independent tumour. Supplementary Fig. 1B-In accordance with journal requirements, 1 kb DNA ladder has been added to Supplementary Fig. 1B to indicate band sizes on the gel. Supplementary Fig. 5D-For the cell death assay dataset, values previously reported as technical replicates have been updated to reflect biological replicate values. This dataset contains the raw and processed data supporting the study investigating the role of immunogenic cell death (ICD) and necroptosis in enhancing immunotherapy responses in triple-negative breast cancer (TNBC). The data were generated using a Brca1⁻/⁻p53⁻/⁻ organoid-derived TNBC mouse model that recapitulates the immune landscapes of basal-like tumours. The dataset includes experimental data assessing tumour response, immune infiltration, and therapeutic efficacy following immune checkpoint blockade (anti–PD-1), IAP antagonism, caspase blocking and STING agonist treatment. Genetic perturbation experiments targeting key necroptosis regulators (including Ripk1 and Mlkl) in tumour and stromal compartments are also represented, enabling mechanistic dissection of tumour-intrinsic and microenvironmental contributions to therapy response. These data demonstrate that RIPK1-driven necroptosis is required for effective ICD and synergistic antitumour immunity, and that coordinated necroptosis in both tumour and stromal cells is critical for durable tumour control and immune memory. Additionally, the dataset captures experiments showing that immunologically “cold” tumours can be sensitised to ICD-based immunotherapy through STING pathway activation.