Clone tuj1

The following primary antibodies were used: Mouse anti-OCT3/4 (Santa Cruz Biotechnology, Clone C-10, #sc-5279, 1:50), Rabbit anti-CDX2 (BioGenex, Clone EP25, #NU777-5UC, 1:100), Mouse anti-CDX2 (BioGenex, Clone CDX2-88, #MU392A-5UC, 1:100), Rabbit anti-EOMES (Abcam, #ab23345, 1:100), Mouse anti-SMA (SIGMA ALDRICH, Clone 1A4, #A2547, 1:200), Mouse anti-TUJ1 (Biolegend, Clone TUJ1, #801201, 1:200), Goat anti-GATA6 (R&D Systems, #AF1700, 1:100), Mouse anti-5mC (Millipore, Clone 33D3, #MABE146, 1:200), Rabbit anti-H3K27me3 (Millipore, #07-449, 1:100) and Rabbit anti-H3K9me3 (Abcam, #ab8898, 1:100). The secondary antibodies used were: Goat anti-Mouse IgG FITC-conjugated (Thermo Fisher Scientific, #62-6511, 1:50), Goat anti-Mouse IgG Alexa Fluor^R 555 (Abcam, #ab150114, 1:1000), Donkey anti-Goat IgG Alexa Fluor^TM 488 (Thermo Fisher Scientific, #A11055, 1:1000), Donkey anti-Mouse IgG Alexa Fluor^TM 555 (Thermo Fisher Scientific, #A31570, 1:1000), Donkey anti-Rabbit IgG Alexa Fluor^TM 555 (Thermo Fisher Scientific, #A31572, 1:1000), and Donkey anti-Rabbit IgG Alexa Fluor^TM 488 (Thermo Fisher Scientific, #A21206, 1:1000).

Ten‐week‐old male C57BL/6J mice (24–26 g each) were anesthetized and mounted in a stereotactic frame for trigeminal ganglion (TG) microinjection using the following coordinates: 4.3 mm rostral, 1.5 mm lateral, and 6.24 mm ventral to the lambda.^[66] 3 µL of each NV formulation was injected to the TG of each mouse (n = 3–5 per group). 3 µL of PBS was injected as a control. After 18 h, the animals were euthanized, and TGs were harvested and analyzed with FACS using mouse antineuron‐specific β‐III‐tubulin‐APC (1:100; Clone TUJ‐1, R&D systems) or Brilliant Violet 421 anti‐GFAP antibody (1:80; Clone 2E1.E9, BioLegend). Study protocols were approved by the University of Texas MD Anderson Cancer Center Institutional Animal Care and Use Committee (IACUC).