Fig. 3: SPI1 inhibits ferroptosis by repressing ACSL4 targets.
From: Therapeutic targeting SPI1 in combination with erastin promotes ferroptosis in ccRCC
A Western blot examined the efficiency of SPI1 knockdown transfected with the shSPI1 in 786-O and ACHN. B qRT-PCR detection of NOX1, SLC7A11, ACSL4, GPX4, HO-1, NRF2 and FTH1 mRNA expression in shCtrl and shSPI1 groups (n = 3). C SPI1and ACSL4 proteins were detected in the si-Ctrl group and si-SPI1 group of 786-O and ACHN cell lines. D It is predicted that SPI1 may bind to the BS1 and BS2 sites of the 2000 bp sequence upstream of the ACSL4 transcription start site according to the HumanTFDB. E The dual-luciferase reporter assay showed the activity of the ACSL4 promoter fragment in 293 T cell line (n = 3). F ChIP-PCR analysis of the binding of SPI1 to the promoter of ACSL4 in 786-O and ACHN cell lines (n = 3). G CCK-8 assays were used to analyse the effect on cell viability in 786-O and ACHN. H, I Flow cytometry was used to detect lipid peroxidation level in 786-O and ACHN (n = 3). J, K MDA and Fe2+ levels were measured separately in 786-O and ACHN (n = 4). Data are presented as the mean ± SD and analyzed by t -test or Kruskal-Wallis test. *: P < 0.05, **: P < 0.01, ***: P < 0.001.
