Ab235166

The western blot assay was carried out as previously described (Zhao et al., 2021 (link)). The primary antibodies against caspase-3 (Abcam, ab65080), cyclin D1 (Abcam, ab226977), CDK4 (Abcam, ab137675), p21 (Abcam, ab227443), FPN (Abcam, ab235166), TF (Abcam, ab84036), FTH1 (Abcam, ab65080), GPX4 (Abcam, ab125066), Nrf2 (Abcam, ab137550), HO-1 (Abcam, ab13243) and β-actin (Abcam, ab8227) were used in this investigation.

Freshly isolated mouse retinas were homogenized and lysed in ice-cold Radioimmunoprecipitation assay (RIPA) buffer (P0013; Beyotime). A Bicinchoninic Acid Kit (Pierce, Rockford, IL, USA) was used to quantify the total protein content. Western blotting was performed as described in our previous study [35 (link)] using the following antibodies: Anti-ACSL4 (as used in IF; 1:3000), anti-GPX4 (as used in IF; 1:1000), anti-FTH1 (as used in IF; 1:1000), anti-divalent metal transporter 1 (DMT1; ab55735; Abcam; 1:1000), anti-transferrin receptor (TR; ab84036; Abcam; 1:1000), anti-ferroportin 1 (Fpn1; ab235166; Abcam; 1:500), anti-nuclear receptor coactivator 4 (NCOA4; sc-373739; Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:500), and anti-β-actin (GB12001; Servicebio; 1:3000). The original western blots for these results are provided in Supplementary File 1.