a, HBEC76 cell pellets (control versus PFKP–GFP overexpression) stained with PFKP antibody used for subsequent immunohistochemistry analysis. The experiment was performed once. b, Microphotograph showing annotated areas of malignant cells (red), tumour stroma (green) and normal bronchial epithelium (yellow) using HALO v.2.3 software in a TMA core of lung-cancer tumour tissue. c, Microphotographs showing the workflow for image analysis using HALO v.2.3 software. From left to right and top to bottom: PFKP staining in a TMA core with lung adenocarcinoma and bronchial epithelium; Halo mark-up image showing the compartments analysed (bronchial epithelium in yellow, tumour stroma in green and malignant cells in red); and Halo mark-up images showing the image analysis using the Halo algorithm to detect cells with PFKP cytoplasmic immunohistochemistry (IHC) expression per bronchial epithelium, tumour stroma and malignant cells. Column charts show frequencies of different levels of expression (0, 1+, 2+ and 3+) of PFKP per tissue type.