Sc 80

Briefly, frozen-sectioned were incubated with primary antibodies at 4 °C overnight after blocking with appropriate sera for 1 h at RT. After washing three times, sections were incubated with the appropriate fluorescent secondary antibody and DAPI and then mounted. Primary antibodies that were used included: rabbit anti-OMA1 monoclonal (PA5-106421, 1:50; Thermo Fisher Scientific), mouse anti-βIII tubulin monoclonal (801202, 1:500, BioLegend, San Diego, CA, USA), mouse anti-AP2α monoclonal (3B5, 1:50; Developmental Studies Hybridoma Bank, Iowa City, IA, USA), rabbit anti-p-eIF2α monoclonal (D9G8, 1:50, Cell Signaling, Danvers, MA, USA), rabbit anti-ATF4 (monoclonal, D4B8, 1:50, Cell Signaling; polyclonal, 10835-1-AP, 1:50, Proteintech, Rosemont, IL, USA), mouse anti-ATF4 monoclonal (SC-390063, 1:50, Santa Cruz Biotechnology, Dallas, TX, USA), guinea pig anti-parvalbumin polyclonal (195004, 1:500, Synaptic System, Göttingen, Germany), mouse anti-CRALBP monoclonal (ab15051, 1:500, Abcam), mouse anti-PKCα monoclonal (SC-80, 1:200, Santa Cruz Biotechnology), and rabbit anti-TOMM20 polyclonal (11802-1-AP, 1:50, Proteintech). Fluorescence signals were quantified using ImageJ.

LPS from Escherichia coli 0111:B4 (L2630), LPS from Salmonella enterica serotype minnesota Re 595 (L9764), peptidoglycan (79682), and actin antibody (A2228) were purchased from Sigma (St. Louis, MO, USA). Antibodies for phospho-PKC-α (sc-12356), phospho-PKC-δ (sc-11770), PKC (sc-80), phospho-IκB-α (sc-8404), IκB-α (sc-371), iNOS (sc-651), COX-2 (sc-1745), ERK1/2 (sc-135990), JNK1 (sc-1648) and p38α (sc-535) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibody for NLRP3 (AG-20B-0006-C100) was purchased from Adipogen (San Diego, CA, USA). Mouse IL-1β (88-7013-88), mouse IL-6 (88-7064-88), mouse TNF-α (88-7324-88), human IL-6 (88-7066-88) and human TNF-α (88-7346-88) ELISA kits were purchased from eBioscience (San Diego, CA, USA). MAPK family antibody sampler kit (9910T) was purchased from Cell Signaling Technology, Inc (Danvers, MA, USA). Pam3CSK4 (tlrl-pms), ATP (tlrl-atp), nigericin (tlrl-nig) and monosodium urate crystal (MSU) (tlrl-msu) were purchased from InvivoGen (San Diego, CA, USA).