Cellsens dimension software v1.12

To investigate proliferation, colony formation ability was assessed. A total of 300 cells (murine and human melanoma cell lines) was seeded in a 6-well plate and cultivated for 10 days (8% CO₂ at 37°C). After washing twice with PBS, cells were fixed and stained with 6.0 glutaraldehyde and 0.5% crystal violet (Sigma-Aldrich; Merck KGaA), respectively, for 1 h at room temperature). The colony (>50 cells) size was analyzed using cellSens dimension-software (V1.12) and an IX83 fluorescence microscope (both Olympus Corporation).

Anterior pectoralis major muscle was fixed in 10% neutral buffered formalin, trimmed, processed through Sakura Tissue-Tek VIP 5, and embedded in paraffin. Tissue sections (4–5 um) were stained with hematoxylin and eosin (H&E) and Masson's Trichrome stains using an adapted chicken breast muscle specific protocol (Vanhatalo et al., 2021 ). Images were captured using a BX43F microscope fitted with a DP80 digital and cellSens Dimension software v.1.12 (Olympus). Muscle histopathology scores were based on macrophage infiltration, tissue damage, adipose cell and collagen presence as follows: Mild (some macrophage infiltration), Moderate (few macrophage phagocytosis and appearance of fibrotic tissue), and Severe (high levels of macrophage infiltration with complete muscle destruction) (Figure S1E–L) (Kuttappan et al., 2013 (link)).