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Figure 3. Inhibition of N-glycosylation reduced CSLC-associated phenotypes and restored paclitaxel sensitivity. (A and B) Kaplan-Meier curves showing the OS (A) and DFS (B) in lung cancer patients with high vs. low expression of a seven-gene N-glycosylation signature (n = 241 per group); (C) Forest plot showing the gene-specific HRs for DFS based on the expression of the seven N-glycosylation-related genes; (D and E) Relative cell growth of A549-TR (D) and H1299-TR (E) cells following 48 h of treatment with 117 nM paclitaxel alone or in combination with TM; n = 3; (F-I) Representative images and quantification of sphere formation in A549-TR (F and G) and H1299-TR (H and I) cells following combined paclitaxel and TM treatment; Scale bar, 100 μm; n = 3; (J) RT-qPCR analysis of stemness-associated genes in A549-TR spheres treated with TM (0.1 or 0.5 μM), n = 3; (K) Forest plot showing gene-specific SMDs of stemness-associated gene expression following TM treatment; (L and M) Representative western blots (L) and quantification (M) of stemness-associated proteins in A549-TR cells treated with TM (0.1 or 0.5 μM) for 48 h. Data were obtained from three independent biological replicates (n = 3); (N) Quantification of CD104^-CD166⁺CD49f^hi CSLC subpopulations in H1299-TR cells after 48 h of TM treatment, n = 3. n.s., not significant; ^*P < 0.05; ^**P < 0.01. CSLC: Cancer stem-like cells; OS: overall survival; DFS: disease-free survival; HRs: hazard ratios; TM: tunicamycin; RT-qPCR: reverse transcription quantitative polymerase chain reaction; SMDs: standardized mean differences; CIs: confidence intervals; n.d.: not detected; SFA: sphere-forming ability; DMSO: dimethyl sulfoxide.