Fig 1: SCAP and SREBP-1 expression in NSCLC and normal lung tissues. Representative IHC images used to measure the differential expression of (A) SREBP-1and (B) SCAP in NSCLC tissues from patients with stage I, II, III, and IV disease and in non-tumor lung tissue samples. (C–E) Quantification of SREBP-1 and SCAP expression in non-tumor and tumor lung tissue of different stages, with in-depth analysis of SREBP-1 nuclear localization in lung tissue. (F,G) Expression and correlation of SREBP-1 and SCAP at the mRNA level in NSCLC samples from the GSE18842 lung cancer microarray dataset compared to the normal lung samples. * p < 0.05, *** p < 0.001, and **** p < 0.0001.
Fig 2: Enhanced SREBP-1/SCAP/FASN signaling is implicated in the reduced sensitivity of NSCLC cells to cisplatin therapy. (A) The staining intensity scores of the cisplatin-sensitive and cisplatin-resistant samples were compared. Graphical representations in boxplot and micrographs (with p values) indicating the SREBP-1, SCAP, and FASN levels in the cisplatin-sensitive and cisplatin-resistant samples. (B) The qRT-PCR analysis was used to measure the expression of SREBP-1/SCAP/FASN. * p < 0.05, and *** p < 0.001.
Fig 3: miRNA expression profiles of NSCLC cells. (A) Heat map and (B) volcano plot of miRNA expression profiles of NSCLC cells. Differentially expressed (DE) miRNAs were identified from the GEO (GSE102286) database containing 179 total lung patient samples using the criteria of |logFC| > 1 and p < 0.05. (C) Intersection analysis of predicted miRNAs targeting SREBP-1 and SCAP. Predictions were based on the ENCORE database. (D) qRT-PCR and (E) Western blot images indicating the effect of the introduction of hsa-miR-497-5p (mimic) and its efficacy of hsa-miR-497-5p in targeting SREBP-1 in H441R and A549R cells. * p < 0.05.
Fig 4: In Western blotting analysis, the precursor and mature SREBP‐2, SP‐1, and SCAP in cells were treated with curcumin at different concentrations. β‐actin was used as an invariant control for equal loading. Representative was shown from three independent experiments. a, The detection of protein expression and the gray value analysis under different concentrations of curcumin treatment at 24 hr and under 50 μM curcumin treatment at 48 hr *, p < .05, versus the untreated control; #, p > .05, versus the untreated control. b, The expression of protein and the gray value analysis under 25 μM curcumin treatment from 0 to 96 hr cultivation. *, p < .05, versus the untreated control; #, p > .05, versus the untreated control
Fig 5: Fatostatin treatment decreased cisplatin resistance and cancer stemness in NSCLC cells. (A) Graphical representation of the effect of 20- to 120-μM cisplatin on A549R and H441R cells left untreated or pretreated with 5 µM fatostatin. (B) Representative Western blot images indicating protein expression of mature (m) SREBP-1 undergoing cleavage in nuclear (Nuc) compartment (top panel), precursor (p) SREBP-1 in cytoplasm (Cyt), and constitutive expression of SCAP, INSIG1, FASN, and HMGCS-1 in whole lysate of A549R and H441R cells either left untreated or treated with 5-µM fatostatin for 24 h. Lamin B1 and GAPDH were loading control of nuclear and whole lysate, respectively. (C) Bar chart showing the comparative expression of SREBP-1, SCAP, FASN and HMGCS-1 mRNA in H441R and A549R cells treatment with/without 5μM Fatostatin. (D) Effect of 2.5- to 10-μM fatostatin on the percentage of CD133+ A549R and H441R cells, as demonstrated by flow cytometry. (E) Images and graphical representations of the effect of 5-μM fatostatin on the number and size of spheres formed by A549R and H441R cells. Data are represented as means ± SDs from assays performed in triplicate at least 3 times. * p < 0.05.
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