Histological Analysis of Lung Tissue

Mice were culled at indicated time points post-microbeads' administration or 14 days post-LLC injection as described (Branco-Price et al., 2012 (link)). Lungs were fixed in 4% paraformaldehyde and embedded in paraffin and cross-sections (7 μm) were cut at 200 μm intervals throughout the lung. Contiguous sections were stained with haematoxylin and eosin (H&E, Sigma Aldrich, UK), Martius Scarlet Blue (MSB, Sigma Aldrich, UK), and for HIF1α (Novus Biologicals, UK), HIF2α (Novus Biologicals, UK), and Mac2 (BioLegend Ltd, UK) as previously described (Evans et al., 2010 (link), 2014a (link),b (link),c (link); Saha et al., 2013 (link)).