Figure 1. CAFs have greater basal autophagic flux than NFs.
(A) Electron microscopy exhibits highly vesicular architecture of CAFs with heterogeneous electron dense and electron poor organelles compared to NFs. Scale bars represent 0.5 μm. Graph depicts percent autophagosomes/fibroblast relative to NF. Autophagosomes were counted in a total of 36 fibroblasts from each group including 4 explants each from HNSCC or cancer-free subjects. Error bars represent ± SEM.
(B) Representative immunoblot of CAFs compared with NFs with and without CQ (20 μM for 6 h) for LC3 protein conversion and p62. Graph depicts percent cumulative density of LC3 levels in CQ treated lanes relative to NF, in 4 explants each of HNSCC or cancer-free subjects. LC3-II levels were normalized to β-tubulin levels. Error bars represent ± SEM.
(C) Representative immunofluorescent of LC3 (green) puncta, Hoechst nuclear stain (blue), comparing NF with CAFs with and without CQ (80 μM for 2 h) (60x magnification). Cumulative results of LC3 puncta per cell counted by a blinded observer of at least 30 cells each of NFs and CAFs. The experiment was repeated 3 times using 3 explants each from HNSCC or cancer-free subjects. Error bars represent ± SEM.