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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Cancer Res. 2017 Sep 28;77(23):6679–6691. doi: 10.1158/0008-5472.CAN-17-1077

Figure 1. CAFs have greater basal autophagic flux than NFs.

Figure 1

(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.