Fig. 3: Supermeres increase lactate release and transfer cetuximab resistance.
a, Heatmap of normalized spectral counts for select proteins and enzymes involved in glycolysis in sEVs, NV fractions, exomeres and supermeres from DiFi cells. b, GSEA analysis of pathways enriched in metabolic enzymes for supermeres versus sEVs (left) and supermeres versus exomeres (right) from DiFi cells. NES, normalized enrichment score; and FDR, false discovery rate. c,d, Immunoblot analysis of select metabolic enzymes and proteins involved in glycolysis in cells and extracellular samples derived from DiFi (c) as well as PANC-1, SC, LM2-4175, MDAM-MB-231 and HREC (d) cells. e, Immunoblot analysis of ENO2 and LDHA in DiFi whole-cell lysate as well as high-resolution density gradient-fractionated sEVs, NV fractions, and exomeres and supermeres. c–e, Equal quantities (30 µg) of protein from each fraction were analysed. f, Lactate release of CC cells treated with PBS (control) or 50 µg ml−1 supermeres derived from CC, SC or CC-CR cells as the mean ± s.e.m. of n = 3 independent treatments. g, Growth analysis of CC colonies in 3D collagen and treated with 50 µg ml−1 supermere derived from CC, SC or CC-CR cells in the presence or absence of cetuximab for 14 d. Colony counts plotted as the mean ± s.e.m. of n = 3 independent samples. h, Representative images of CC colonies from g. i, Representative low (top) and high (bottom) magnification images of CC colonies treated with SC supermeres. h,i, Scale bars, 200 µm. j, Growth analysis of DiFi colonies in 3D collagen and treated with 50 µg ml−1 sEV-Ps, exomeres and supermeres derived from DiFi cells in the presence or absence of cetuximab for 14 d. Colony counts plotted as the mean ± s.e.m. of n = 6 independent experiments. f,g,j, *P < 0.01, **P < 0.001; two-tailed Student’s t-test. Exom, exomere; super, supermere; WCL, whole-cell lysate; CTL, control; and CTX, cetuximab.
