Tulp3

We performed Western blotting according to a previous study [25 (link)]. In short, cells were lysed using RIPA reagent (Beyotime Biotechnology, Shanghai, China). The protein was quantified using the bicinchoninic acid method (Beyotime Biotechnology). The protein was run on 8% sodium dodecyl sulfate,sodium salt polyacrylamide gelelectrophoresis and then was transferred to polyvinylidene fluoride membrane (Promega). Then, the membrane was blocked by nonfat milk and incubated with primary antibody (BCL-2, #3498, 1:1000, CST; Bax, #50599-2, 1:5000, Proteintech; BCL-XL, #2764, 1:1000, CST; TULP3, #13637, 1:1000, Proteintech; cytochrome C, #ab90529, 1:1000, Abcam, Cambridge, MA, USA; and cleaved caspase-9, #ab2324, 1:1000, Abcam). The membrane was washed twice and incubated with a secondary antibody for 2 h (1:1000, Forevergen, Guangzhou, China). The membrane was washed thrice. The membrane was observed using ECL (Forevergen).

As shown in the research [23 (link)], IHC was performed following standard protocol. The primary antibodies were as follows: BCL-2 (#3498, 1:100; CST, Danvers, MA,USA), Bax (#50599-2, 1:500; Proteintech, Wuhan, China), BCL-XL (#2764, 1:1200; CST), and TULP3 (#13637, 1:500; Proteintech).