Fig. 2

Functional validation of in vitro-derived porcine PSCdMs. A Composite bright-field and fluorescent image showing phagocytosed pHrodo-Red beads fluorescing within porcine PSCdMs. Image taken 3 h after pHrodo bead addition. B Quantification of phagocytosis activity in PAMs (blue) and PSCdMs (red). Graph shows the level of pHrodo bead fluorescence between 0 and 8 h. Mean and SD of two PSCdM and one PAM line from three experiments. C Flow cytometry analysis of PAMs (blue) and PSCdMs (red) 8 h after pHrodo-Red bead addition relative to negative control cells (grey). D RT-qPCR analysis comparing DDX58 and IFNB expression in PAMs and PSCdMs following 4 h pre-treatment with 200 ng/ml LPS or 25 μg/ml poly(I:C) relative to untreated controls. Mean and SD of three experimental replicates. E Confocal Z-stack projected image of PSCdMs 1 h post-infection with EGFP-labelled Salmonella typhimurium. DNA is stained with DAPI (blue) and actin filaments with phalloidin (red). F Ratio of colony-forming Salmonella typhimurium recovered from infected PAMs and PSCdMs at 3 h post-infection relative to T0. Mean and SD of duplicate plates from two dilutions. G Confocal image of PSCdMs 24 h post-infection with EGFP-labelled Toxoplasma gondii. DNA is stained with DAPI (blue) and actin filaments with phalloidin (red). H RT-qPCR analysis of CCL22 and IRF7 expression in uninfected and Toxoplasma gondii-infected PAMs and PSCdMs. Mean and SD of three experimental replicates