Fig 1: The correlations of ACSL4 and GPX4 density with tumor size, ER, PR, and Ki-67. Notes: Distribution of the data is present along the diagonals. The upper part exhibits correlation coefficients and P values calculated by Spearman's rank correlation analysis (* P < 0.05, ** P < 0.01, and *** P < 0.001). The lower part shows scatter plots with smooth fitting curves. There was a significant correlation between GPX4 and Ki-67 (r=-0.16, P=0.026), ER and PR (r=0.67, P < 0.001), ER and Ki-67 (r=-0.35, P < 0.001), and PR and Ki-67 (r=-0.28, P < 0.001). Abbreviations: ER, estrogen receptor; PR, progesterone receptor.
Fig 2: Subgroup analysis of pCR according to the combination of ACSL4 and GPX4 expression levels. Abbreviations: pCR, pathological complete response; OR, odds ratio; CI, confidence interval; T, tumor; HorR, hormone receptor; HER2, human epidermal growth factor receptor 2; BMI, body mass index.
Fig 3: Chmp1a knockdown enhanced ferroptosis through iron accumulation.a (left) Representative BODIPY 581/591 C11 staining of siControl and siChmp1a transfected cell following sham or cisplatin treatment. Scale bar: 50 µm. (right) Quantification of the oxidized vs. reduced probe (n = 5). b Relative mRNA level of Acsl4 in the kidneys of folic acid and cisplatin-treated wild-type and Chmp1a+/- mice. Sham-treated group: WT (n = 3), Chmp1a+/- (n = 3); FA-treated group: WT (n = 5), Chmp1a+/- (n = 5); cisplatin-treated group: WT (n = 4), Chmp1a+/- (n = 5). c Western blots of ACSL4 in kidneys of folic acid-treated wild-type and Chmp1a+/- mice. d Representative images of ACSL4 immunostaining of kidney sections from folic acid-treated wild-type and Chmp1a+/- mice. Scale bar: 10 µm. e LDH level of siControl and siChmp1a transfected cell with or without cisplatin and ferroptosis inhibitor liproxstatin1 (n = 3). f (upper) Western blots of CD63 of kidney exosomes of wild-type and Chmp1a+/- mice. (bottom) Quantification of three independent experiments (n = 3). g Ferrous iron concentrations (normalized to kidney weight) of kidneys of cisplatin-treated wild-type and Chmp1a+/- mice. Sham-treated group: WT (n = 3), Chmp1a+/- (n = 3); cisplatin-treated group: WT (n = 4), Chmp1a+/- (n = 5). h Serum creatinine level of control and Chmp1a+/- mice with or without cisplatin and liproxstatin injection. Sham-treated group: WT (n = 3), Chmp1a+/- (n = 3); cisplatin-treated group: WT (n = 5), Chmp1a+/- (n = 5); cisplatin and liproxstatin-treated group: WT (n = 4), Chmp1a+/- (n = 4). i Relative transcript level of injury marker Kim1 in kidneys of control and Chmp1a+/- mice with or without cisplatin and liproxstatin injection. Sham-treated group: WT (n = 3), Chmp1a+/- (n = 3); cisplatin-treated group: WT (n = 5), Chmp1a+/- (n = 5); cisplatin and liproxstatin-treated group: WT (n = 4), Chmp1a+/- (n = 4). All data are represented as mean ± SEM. P value was calculated by two-way ANOVA with post hoc Tukey test for a, b, e, g, h, i. P value was calculated by two-tailed t-test for f. P < 0.05 is statistically significant. A Source Data file is available for this figure.
Fig 4: ACSL4 depletion suppresses the growth of HCC cells and c-Myc protein level in vivo.a Representative images of tumor size in nude mice injected subcutaneously with Huh7 cells infected with either shNC or shACSL4 (n = 6, for each experimental group). The mice were killed on the 28th day for tumor weight analysis. b Tumor growth rates and tumor weight in Huh7-shNC group and Huh7-shACSL4 group. c Representative images of tumor size in nude mice injected subcutaneously with SMMC-7721 cells infected with either shNC or shACSL4. The mice were killed on the 28th day for tumor weight analysis (n = 6, for each experimental group). d Tumor growth rates and tumor weight in SMMC-7721-shNC group and SMMC-7721-shACSL4 group. The data shown are the mean ± SD of tumor volume and tumor weight from six nude mice per group (***p < 0.001). The data were analyzed using Student’s t-test. e ACSL4 and c-Myc protein levels in Huh7 indicated xenografts were detected by western blot. f H&E staining and the expression of ACSL4 and c-Myc in tumor sections by IHC analysis (scale bar: 100 µm; magnification: ×200).
Fig 5: ACSL4 promotes the proliferation of HCC cells in vitro.a Effect of ACSL4 depletion on the proliferation of Huh7 and Hep3B cells by CCK-8 assay. b Photos for colony formation (left) and bar graph (right) in ACSL4-depleted Huh7 and Hep3B cells. c Effect of ACSL4 overexpression on the proliferation of Bel-7402 and PLC/PRF5 cells by CCK-8 assay. d Photos for colony formation (left) and bar graph (right) in ACSL4 overexpressed Bel-7402 and PLC/PRF5 cells. e Effect of ACSL4 depletion or overexpression on cell-cycle distribution in HCC cells by FACS. f Effect of ACSL4 depletion or overexpression on G1/S cell-cycle genes in HCC cells by western blotting. GAPDH was used as a loading control. Data are from three independent experiments and expressed as mean ± SD. **p < 0.01, ***p < 0.001. The data were analyzed using Student’s t-test.
Supplier Page from Abcam for Anti-FACL4 antibody [EPR8640]