Supplementary Fig. 1: Markers and major cell types identified by the t-CyCIF immune panel.

a) Canonical immune cell types and their markers. Seven major immune cell subtypes were assayed using a panel of 16 markers. Three additional markers (Ki-67, α-SMA and pan-keratin) were used to identify cell states (Ki-67 for proliferative cells) or separate immune cells from tumour cells (keratin-positive) or stromal cells (α-SMA-positive). b) Actual immune subpopulation identified from t-CyCIF immune profiling of LUNG-3-PR. 23,079 immune cells (keratin/α-SMA-negative cells) from the sample were used for binary gating of 15 different markers. A total of 1,356 different subpopulations were identified, of which 37 subpopulations represented >0.5% of total immune cells. The four subpopulations highlighted here are CD45⁺/IBA1⁺ (macrophage or dendritic cells), CD45⁺/CD20⁺ (B cells), CD45⁺/CD3⁺/CD8a⁺ (cytotoxic T cells) and CD45⁺/CD3⁺/CD4⁺ (helper T cells). Asterisks label 19 common immune cell subtypes.