serum free medium  (R&D Systems)

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Inhibiting FAT1 Blocks Metabolic Bypass to Enhance Antitumor Efficacy of TCA Cycle Inhibition through Suppressing CPT1A-Dependent Fatty Acid Oxidation.

doi: 10.1002/advs.202502146

Figure Lengend Snippet: Figure 1. Mutant FAT1 functions as a tumor promoter in HNSCC. A) Top 10 most frequently mutated genes in TCGA HNSCC cohort (n = 523). B) Frequency of FAT1 mutation in HPV- and HPV+ tumor subtypes in TCGA HNSCC cohort. The number and percentage of cases with mutations are displayed. NA, not annotated. C) Clinical evaluation of FAT1 mutations in HPV- tumors in TCGA HNSCC cohort. Points represent the individual score of each variable at different values, and total points represent the total score of the corresponding individual scores for all variables. Pr (prediction) represents the predicted overall survival (OS) at indicated years. D) Calibration curve assessing the reliability of nomogram model for OS in HPV- HNSCC patients with FAT1 mutations. Calibration curve values greater than 0.75 suggest the credibility of the nomogram model. E) High TMB associated with FAT1 mutations as a potential risk factor for OS in HPV- HNSCC patients with FAT1 mutations in TCGA cohort. F) FAT1 KO efficiency in SCC1 and Tu686 cells determined by Western blot. G–I) Effect of FAT1 KO on cell proliferation (for 3 days), colony formation (for 14 days), and tumorsphere formation (for 14 days) in SCC1 and Tu686 cells. In (H) and (I), the experiments were performed in triplicate and quantitative data are shown in the right panel. (J, K) Representative flank tumors and tumor growth curve J), and tumor weight K) in the indicated treatment groups. FAT1 KO or parental SCC1 cells were implanted to the right flank of 6-week-old NSG mice (n = 5/group), and tumor size was measured by a digital caliper weekly for 35 days. Bars express mean ± SD. Statistical analyses were conducted using unpaired two-tailed Student’s t-test. *p < 0.05; **p < 0.01.

Article Snippet: Tumorsphere Formation Assays: Dissociated single HNSCC cells with or without FAT1 KO were plated on Corning Costar 6-well ultra-low attachment plates (cat# 3471, In Vitro Technologies) at a density of 1 × 104 cells per mL and grown in StemXVivo Serum-free Tumorsphere media (cat# CCM012, R&D Systems, In Vitro Technologies).

Techniques: Mutagenesis, Western Blot, Two Tailed Test

Journal: Advanced Science

Article Title: Inhibiting FAT1 Blocks Metabolic Bypass to Enhance Antitumor Efficacy of TCA Cycle Inhibition through Suppressing CPT1A‐Dependent Fatty Acid Oxidation

doi: 10.1002/advs.202502146

Figure Lengend Snippet: Mutant FAT1 functions as a tumor promoter in HNSCC. A) Top 10 most frequently mutated genes in TCGA HNSCC cohort (n = 523). B) Frequency of FAT1 mutation in HPV‐ and HPV+ tumor subtypes in TCGA HNSCC cohort. The number and percentage of cases with mutations are displayed. NA, not annotated. C) Clinical evaluation of FAT1 mutations in HPV‐ tumors in TCGA HNSCC cohort. Points represent the individual score of each variable at different values, and total points represent the total score of the corresponding individual scores for all variables. Pr (prediction) represents the predicted overall survival (OS) at indicated years. D) Calibration curve assessing the reliability of nomogram model for OS in HPV‐ HNSCC patients with FAT1 mutations. Calibration curve values greater than 0.75 suggest the credibility of the nomogram model. E) High TMB associated with FAT1 mutations as a potential risk factor for OS in HPV‐ HNSCC patients with FAT1 mutations in TCGA cohort. F) FAT1 KO efficiency in SCC1 and Tu686 cells determined by Western blot. G–I) Effect of FAT1 KO on cell proliferation (for 3 days), colony formation (for 14 days), and tumorsphere formation (for 14 days) in SCC1 and Tu686 cells. In (H) and (I), the experiments were performed in triplicate and quantitative data are shown in the right panel. (J, K) Representative flank tumors and tumor growth curve J), and tumor weight K) in the indicated treatment groups. FAT1 KO or parental SCC1 cells were implanted to the right flank of 6‐week‐old NSG mice (n = 5/group), and tumor size was measured by a digital caliper weekly for 35 days. Bars express mean ± SD. Statistical analyses were conducted using unpaired two‐tailed Student's t ‐test. * p < 0.05; ** p < 0.01.

Article Snippet: Dissociated single HNSCC cells with or without FAT1 KO were plated on Corning Costar 6‐well ultra‐low attachment plates (cat# 3471, In Vitro Technologies) at a density of 1 × 10 4 cells per mL and grown in StemXVivo Serum‐free Tumorsphere media (cat# CCM012, R&D Systems, In Vitro Technologies).

Techniques: Mutagenesis, Western Blot, Two Tailed Test