Fig. 2

NAT10 KD impairs tumor migration, invasion, and PNI in PDAC. (A) Representative immunoblots (left) and quantification (right) of NAT10 in human pancreatic ductal epithelial (hTERT-HPNE) and human PDAC cell lines. (B-C) Relative mRNA expression (B) and representative immunoblots (C) of NAT10 in NAT10 KD (via shRNA) and control PANC-1 or MIA PaCa-2 cells. (D-E) Representative image and quantification of Transwell migration and invasion in NAT10 KD and control PANC-1 (D) or MIA PaCa-2 (E) cells. Scale bar: 200 μm. (F-G) PANC-1 (F) or MIA PaCa-2 (G) cells were cocultured with DRGs into Matrigel. Representative images (left) on Day 3 and 6 are shown. The arrowhead indicates tumor cell invasions. Quantification (right) of relative recruited cancer cells on Day 6 is shown. Scale bar: 200 μm. (H) NAT10 KD or control PANC-1 cells were injected into the sciatic nerve of SCID mice (n = 4). Representative immunofluorescence image of the Schwann cell marker S100β and tumor marker CK19 reveals nerve invasion of NAT10-KD or control PANC-1 cells. Scale bar: 100 μm. (I-J) Nerve function score (hind limb function, I) and sciatic nerve function index (hind paw span, J) were measured every 7 days. *p < 0.05, **p < 0.01, ***p < 0.001. (A-B), and (D-E), One-way ANOVA. (F-G), Student’s t-test