Anti pex5

Anti-PEX5 was purchased from GeneTex (Irvine, CA, USA). Anti-catalase, anti-LAMP-1, anti-PCNA, anti-cathepsin B, and anti-cathepsin D were bought from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-LAMP-2 and anti-LC3 were obtained from Abcam (Cambridge, MA, USA). Anti-TFEB was purchased from MyBioSource (San Diego, CA, USA). Anti-TFEB, anti-p-p70S6K, anti-p70S6K, anti-p-S6R, anti-S6R, anti-p-4E-BP-1, anti-4E-BP-1, and anti-TSC2 were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-PMP70 was bought from Thermo Fisher Scientific (Waltham, MA, USA). Anti-ACOX1 was purchased from Proteintech (Chicago, IL, USA). Anti-DBP (HSD17B4) was bought from OriGene Technologies (Rockville, MD, USA). Anti- SQSTM1/p62 was obtained from Sigma-Aldrich (St. Louis, MO, USA).

All plasmids in this study were constructed through polymerase chain reaction (PCR)-base strategy and sequenced to confirm their identity. Full cDNA sequence of mouse Hsd17b4 was purchased from Darmacon (USA) (MMM4769-202765308). Full sequence Hsd17b4 was constructed in the pEBB-Flag and pEBB-GFP vector (Min et al., 2020 (link)). The untagged protein expressing construct was generated as well. In order to identify PS binding region of Hsd17b4, the three domains of Hsd17b4 were constructed in the pEBG-GST vector. GST-Hsd17b4^1-305, GST-Hsd17b4^321-621, and GST-Hsd17b4^633-730 contain the hydroxyacyl-CoA dehydrogenase domain, Enoyl-CoA hydratase 2 domain, and the SCP2-like domain of Hsd17b4. HA-Tim-4^IgV was previously reported (Lee et al., 2019 (link)).
The antibodies used in this study were anti-Hsd17b4 (15116-1-AP [Proteintech, USA] and NBP2-46005 [Novus, USA]), anti-GST (SC-138; Santa Cruz Biotechnology, USA), anti-Catalase (ab209211; Abcam, UK), anti-Actin (SC-47778; Santa Cruz Biotechnology), anti-Pex5 (GTX109798; GeneTex, USA), anti-Pmp70 (ab3421; Abcam), anti-HA (sc-7392; Santa Cruz Biotechnology), Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488, and Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 (A11008 and A21422; Thermo Fisher Scientific, USA).