Rabbit anti active caspase 3

Rabbit anti-KCC2 pan polyclonal antibodies [32 (link)] were produced and purified by Innovagen AB and used at a dilution of 1:1000 for IHC. Primary antibodies Mouse anti-NeuN, clone A60 (Merck Millipore, MAB377, 1:300), mouse anti-GFAP (Merck Millipore, MAB360, 1:500), rabbit anti-Iba1 (Wako, 019-19741, 1:500), and rabbit anti-active® caspase 3 (Promega, G7481, 1:200) were used. The NGFR p75 (MC-192) mouse antibody was from Santa Cruz and was used 1:1000 with the blocking solution from Mouse on Mouse (M.O.M.™) Basic Kit (Vector Laboratories). Secondary antibodies, goat anti-rabbit, or anti-mouse conjugated to Alexa Fluor® 488, 555, 568, or 647 were used at dilutions of 1:500. p75^NTR KO mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA) [33 (link)].

Rabbit polyclonal antibodies against individual LRRC8 subunits had been generated in our laboratory and their specificity had been confirmed in western blots using KO cell lines as controls as described^{25 (link),29 (link),30 (link)}. The following commercial antibodies were used: guinea pig anti-insulin (Dako, A0564; 1:500), mouse anti-glucagon (Abcam, ab10988; 1:1000), mouse anti-β-actin (Sigma, A2228; 1:2000), rabbit anti-RFP (Rockland, 600401379; 1:1000), rabbit anti-Kir6.2 (Abgent, AG1143; 1:200), goat anti-NLRP3 (Lifespan Biosciences, LS-B1766; 1:200), rabbit anti-active caspase 3 (Promega; G748A1; 1:400). Secondary antibodies were from Molecular Probes (coupled to Alexa-488 or Alexa-555 or Alexa 633), or from Jackson ImmunoResearch (coupled to horseradish peroxidase). Quantification of western blots used ImageJ. LRRC8A levels were normalized to actin controls on the same blot. Uncropped western blots are shown in Supplementary Fig. 7.