Fig. 2

Comparison of bone marrow derived macrophages (BMDMs) isolated from young and old mice

(a) Experimental timeline. Bone marrow-derived cells (BMCs) were isolated, differentiated with M-CSF, washed and switched to EV-depleted FBS medium at D6, and analysed at D8. (b) Representative flow cytometry histograms of young (Y_) and old (O_) mouse BMCs at D1 and BMDMs at D8. Positive % for F4/80 (Y axis) and CD11b (X axis) is shown. N ≥ 3 independent batches were analysed. The grey histogram shows isotype control samples. (c) Flow cytometry showing positive population (X axis) for CD206, F4/*), CD11b and CD11c. N = 3 independent batches were analysed. Samples were compared by two-way ANOVA. (d) Quantification of macrophage markers by RT-qPCR in naïve Y_ (n = 9) and O_ BMDMs (n = 3) at D8. Expression was normalised to Hnrnpa1 and Y_BMDMs and O_BMDMs were compared by one-way ANOVA, with Tukey’s post-test, indicated by *. (e) Example images of Y_/O_BMDMs in naïve, IL-4 or LPS-polarised conditions. Data points show samples from separate mice. Bar heights show the mean, error bars show the standard error of the mean. * = P ≤ 0.05, ** = P < 0.01, *** = P ≤ 0.001, **** = P ≤ 0.0001