Ultrasensitive sp immunodetection kit

For immunohistochemical staining, normal and 3-day infected eyes from C57BL/6 and BALB/c mice were enucleated (n = 3/group/time), immersed using ice-cold PBS, embedded in Tissue-Tek OCT compound (Miles, Elkhart, IN, USA), and frozen in liquid nitrogen. Immunohistochemical staining was performed with the UltraSensitive SP Immunodetection Kit (Maixin, Inc., Fuzhou, China) following the manufacturer’s protocol. Primary antibodies (rabbit anti-mouse STING) were purchased from PeproTech (Rocky Hill, NJ, USA). Controls were similarly treated, but the primary antibody was replaced with isotype-matched goat IgG. For immunofluorescent staining, cells were seeded on sterile glass cover slips, cultured overnight, and then fixed with 4% paraform (Sigma) in 4°C. Slips were sequentially incubated with rabbit anti-mouse STING Ab (1:200, PeproTech), rabbit anti-mouse NF-κB (1:200, Cell signaling), and Alexa Fluor 488-conjugated goat anti-rabbit IgG Ab (1:1,000, Millipore, Billerica, MA, USA), followed by incubation with 4,6-diamino-2-phenyl indole (1:10,000, Sigma) for nuclear staining. Controls were similarly treated, but the primary Ab was replaced with isotype-matched IgG. For histopathology, sections were HE stained as described by others (29 (link)). All sections were visualized with a Carl Zeiss microscope (Carl Zeiss, Inc., Oberkochen, Germany).

Infected eyes were enucleated (n = 3/group/time) at 1 and 5 days p.i. from WT and Trem2^−/− B6 mice, embedded in Tissue-Tek OCT compound (Miles, Elkhart, IN, USA) and frozen in liquid nitrogen. 8 µm thick sections were cut and mounted to glass slides. For histopathology, sections were hematoxylin-eosin (HE) stained as described by others (25 (link)). Immunohistochemical staining was performed with the UltraSensitive SP Immunodetection Kit (Maixin, Inc., Fuzhou, China) according to the manufacturer’s instructions. Primary antibodies rabbit anti-mouse capase-1 (p10) (clone: M-20; Cat: sc-514) were purchased from Santa Cruz. All sections were visualized with a Carl Zeiss microscope (Carl Zeiss Inc.).