Skip to main content
. 2022 Aug 10;78(6):617–630. doi: 10.1136/thoraxjnl-2021-218194

Figure 6.

Figure 6

Transfer of miR181a is critical for the immunomodulatory effects of MSC-EVs in vivo. (A) Total protein concentrations in the BALF samples 24 hours after LPS administration (n=5–7 mice per group). (B) Total leucocyte counts (left graph) and neutrophil cell counts (right graph) in the BALF samples (total cells (n=5–7 mice per group); neutrophils (n=2–4 mice per group)). (C) Representative images of BALF cytospin preparations demonstrating cell recruitment to the airspaces 24 hours after LPS administration. Images were taken using Leica Epifluorescence DM5500 microscope (Objective lens ×20). (D) BALF levels of TNF-α and keratinocyte-derived chemokine (KC, murine analogue of interleukin-8) (n=5–7 mice per group). (E) Representative confocal microscopy of alveolar macrophages, isolated from BALF 24 hours after LPS administration and stained with anti-CD68 (alveolar macrophage marker) and anti-pSTAT5 Ab. Arrows indicate co-localisation of CD68 and pSTAT5. The images were taken using Leica SP8 confocal microscope with a 100× oil-immersion objective (n=3, scale bar=50 µm). Quantitative fluorescence intensity was analysed by Image J software (MFI-pSTAT5). (F) Representative confocal microscopy of alveolar macrophages, isolated from BALF samples 24 hours after LPS administration and stained with anti-CD68 and anti-SOCS1 Ab. Arrows indicate co-localisation of CD68 and SOCS1. Images were taken using Leica SP8 confocal microscope with a 100× oil-immersion objective (scale bar=50 µm). Quantitative fluorescence intensity was analyses by Image J software (MFI-SOCS1). Data are represented as mean±SD. Kruskal-Wallis test with post hoc Dunn’s test (A, B, D, E, F). Ab, antibody; BALF, bronchoalveolar lavage fluid; DAPI, 4',6-diamidino-2-phenylindole; EVs, extracellular vesicles; LNA, locked nucleic acid; LPS, lipopolysaccharide; MFI, mean fluorescent intensity; MSCs, mesenchymal stromal cells; PBS, phosphate-buffered saline; pSTAT, phosphorylated STAT; SOCS1, suppressor of cytokine signalling 1; STAT, signal transducers and activators of transcription ;TNF, tumour necrosis factor.