Fig. 7.

FSP1 blockade is crucial for ferroptosis vulnerability in HCC. (A) CCK-8 assays of HDLBP silenced HCC cells transfected with control vector (Vector) and LncFAL expression vector (LncFAL) following treatment with erastin (1–20 μM) for 24 h. (B) Calcein-AM/PI staining of HDLBP silenced HCC cells transfected with Vector and lncFAL following treatment with erastin (4 μM) for 24 h. Right panel: relative quantitative analysis of Calcein-AM/PI staining. C-AM, Calcein-AM. Scale bars = 200 μm. (C) CCK-8 assays of lncFAL-overexpressing HCC cells transfected with FSP-si or CTL-si following treatment with erastin (1–20 μM) for 24 h. (D) Calcein-AM/PI staining of lncFAL-overexpressing HCC cells transfected with FSP-si or CTL-si following treatment with erastin (4 μM) and with/without Fer-1 (2 μM) for 24 h. (E) CCK-8 assays of lncFAL-overexpressing HCC cells transfected with FSP1-si following treatment with erastin (1–20 μM) and with/without Fer-1 (2 μM) for 24 h. (F) CCK-8 assays of HDLBP silenced HCC cells treated with erastin (1–20 μM) and with/without ubiquinol (1 μg) for 24 h. (G) CCK-8 assays of lncFAL silenced HCC cells treated with erastin (1–20 μM) and with/without ubiquinol (1 μg) for 24 h. (H) Representative images of the xenograft, tumour volume and tumour weight 28 days after inoculation of lncFAL-overexpressing PLC5 cells infected with FSP-sh or CTL-sh (n = 5 per group). The tumour volumes were measured every 7 days. (I) Representative images of HE and IHC of Ki67, GPX4, and FSP1 expression in the implanted HCC tumours in each group. Scale bars = 50 μm. The experiments in A-G were independently performed in triplicate (N ≥ 3). The data are presented as the means ± SDs. The statistical analyses were performed by the two-tailed unpaired Student's t-test (A–H).