Fig. 3: Multi-omics analysis reveals significant enrichment in some important signaling pathways and cellular processes in rCAFs compared to sCAFs.
a Heatmap analysis of the transcriptomics data from three different NACT-sensitive and -resistant HNSCC patients derived CAFs respectively. b, c KEGG and GO analysis of the transcriptomics data between sCAFs and rCAFs. Red color highlights the signaling, cellular and biological pathways and molecular functions related to this study. d Volcano plot showing the comparative proteomics profiling analysis of three different NACT-sensitive and -resistant HNSCC patients derived CAFs. e, f KEGG and GO analysis of the proteomics data from three different sCAFs and rCAFs respectively. Red color highlights the signaling, cellular and biological pathways and molecular functions involved in this study. g, h Western blot analysis of the indicated proteins in sCAFs and rCAFs in the presence or absence of 8 μM DDP. Molecular markers (kDa) are shown on the right-hand side. i Annexin V-PI apoptotic assays of rCAFs and sCAFs after treated with or without 8 μM DDP or 15 μM 5’FU. The percentage of viable cells, early apoptotic cells, and late apoptotic cells was determined based on the lower quadrant (Q4), lower right quadrant (Q3), and upper quadrant (Q2), respectively. The red number indicates the percentage of apoptotic cells in each group. j DDP or 5’FU IC50 experiments of sCAFs and rCAFs after treated with or without 10 μM AKT (Perifosine) or 20 μM p65 inhibitors (Maslinic acid) (n = 3 independent experiments). k Representative co-immunofluorescent staining of p-P13K/p-AKT and α-SMA in tumor sections from each group. Bar charts represents the staining intensity in each group as indicated (n = 20 patients, our cohort). Arrow indicates p-P13K or p-AKT and α-SMA double positive cells. Data are shown as means ± S.E.M. **P < 0.01, ***P < 0.001. a–f, k Student’s t test. Scale bars in (k) represent 100 μm.
