Comprehensive Western Blot Analysis of Neural Markers

Western blotting was performed as previously described (Huang et al. 2018 (link)) with the diluted primary antibodies NeuN (1: 300, cat. # ab104224, Abcam), GFAP (1: 300, cat. # ab7260, Abcam), Iba-1 (1: 300, cat. # ab178846, Abcam), caspase-1 (1: 100, cat. # sc-56036, Santa Cruz), NLRP3 (1: 1,000, cat. # DF7438, Affinity), ASC (1: 1,000, cat. # ab47092, Abcam), Gasdermin D (GSDMD, 1: 500, cat. # sc-393581, Santa Cruz), IL-1β (1: 1,000, cat. # ab254360, Abcam), IL-18 (1: 1000, cat. # ab207323, Abcam), BDNF (1: 1000, cat. # ab108319, Abcam), PSD95 (1: 1000, cat. # ab238135, Abcam), Syna (1: 1000, cat. # ab32127, Abcam), and GAP43 (1: 1000, cat. # ab75810, Abcam) and incubated for 12 h at 4 °C. After being washed three times with 0.1 M PBS, the membranes were incubated with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (diluted to 1: 5000, Abcam, USA) for 2 h at RT. Finally, the protein bands were visualized using a Western blotting detection kit (Wanleibio, China).