Fig. 5: Differentiating between Actin and Golgi responses at different effective compound concentrations using cytochalasin D.

A Representative images (Actin and Golgi channels) showing activities of cytochalasin D on Actin- and Golgi-related features at two non-cytotoxic concentrations compared to the DMSO solvent control across four different cell lines. N_Biol = 4. Scale bars = 20 µm. B Corresponding BMC accumulation and magnitude plots showing cytochalasin D effects on feature categories and single features across four different cell lines in CPP, and MCF7 in CP as described in Fig. 3D. C Profile similarity plots showing the correlation of the phenotypic profiles (Spearman correlation of robust z-scores at the feature level, excluding Lyso features) of all reference compounds at each highest non-cytotoxic concentration in MCF-7 cells. Compounds are assigned to one of six clusters (grey-shaded boxes) based on hierarchical clustering. Compounds are color-coded (cyan, orange, red) according to their annotated Actin-, Mito-, or Lyso-related MoA (see Fig. 2A). Colored, dashed boxes highlight correlation scores of compounds with the same annotated MoA. Input feature data (median of all N_Tech = 3 and N_Biol = 4 for each feature) are extracted from CPP (CPP/Harmony, CPP/Cell Profiler) or CP (CP/Harmony) images using Harmony or Cell Profiler image analysis software. Correlation scores of negative control compounds (i.e., saccharine, sorbitol) are shown slightly transparent.