Fig. 5: Silencing of Smad3 enhances the antitumor activity of TANs in vivo.
From: Smad3 is essential for polarization of tumor-associated neutrophils in non-small cell lung carcinoma
A, B Smad3-WT and Smad3-KO mice were treated with anti-Ly6G or control IgG beginning 3 days before inoculation with LLC cells and killed on day 16 and analysed for; A tumor size, and B tumor weight (*P < 0.05, ***P < 0.001 vs. IgG-treated Smad3-WT mice; ##P < 0.01, ###P < 0.001 vs. IgG-treated Smad3-KO mice; one-way ANOVA). C–F Adoptive transfer of Smad3-knockout (KO-BMDN), but not wild-type BMDNs (WT-BMDN), on days 15 and 20 significantly inhibited the growth of LLC tumors in Smad3-WT mice as shown by: C tumor size, and D tumor weight, on day 25. E Immunohistochemistry and H&E staining shows that adoptive transfer of WT-BMDNs increases the proportion of neutrophils within the TME in LLC tumors, while the transfer of KO-BMDN resulted in an even greater proportion of neutrophils in the TME. F Immunostaining shows that the transfer of WT-BMDN did not affect the proportions of N1 and N2 TANs in the TME, whereas the transfer of KO-BMDN caused a substantial change in the proportion of N1 and N2 TANs. (*P < 0.05, **P < 0.01, ***P < 0.001 vs. Control, ##P < 0.01, ###P < 0.001 vs. WT-BMDN, one-way ANOVA). Scale bars, 50 μm. A–F Data represents mean ± SEM of 4 mice/group. The exact P values of IgG- vs. anti-Ly6G-treatment are 5A. P = 0.0001 (IgG KO vs. IgG WT, D14), P = 0.0122 (anti-Ly6G KO vs. IgG WT, D14), P = 0.0049 (anti-Ly6G KO vs. IgG KO, D14), P = 0.0001 (IgG KO vs. IgG WT, D16), P = 0. 0001 (anti-Ly6G KO vs IgG WT, D16), P = 0.0085 (anti-Ly6G KO vs. IgG KO, D16), 5B. P = 0.0001 (IgG KO vs IgG WT), P = 0.0001 (anti-Ly6G KO vs. IgG WT), and P = 0.0003 (anti-Ly6G KO vs. IgG KO). Exact P values of BMDN adoptive transfer are 5C. P = 0.0001 (KO-BMDN vs. WT-BMDN, D20), P = 0.0001 (KO-BMDN vs. Control, D20), P = 0.007 (KO-BMDN vs. WT-BMDN, D25), P = 0.0009 (KO-BMDN vs. Control, D25). 5D. P = 0.0068 (KO-BMDN vs. WT-BMDN), P = 0.0014 (KO-BMDN vs. Control), 5E. P = 0.0013 (KO-BMDN vs. WT-BMDN, Ly6G), P = 0.0001 (KO-BMDN vs. Control, Ly6G), P = 0.0226 (WT-BMDN vs. Control, Ly6G), P = 0.0018 (KO-BMDN vs. WT-BMDN,H&E), P = 0.0001 (KO-BMDN vs. Control, H&E), P = 0.0001 (WT-BMDN vs. Control, H&E). 5F. P = 0.0001 (KO-BMDN vs. WT-BMDN,N1 TAN), P = 0.0001 (KO-BMDN vs. Control, N1 TAN), P = 0.0205 (WT-BMDN vs. Control, N1 TAN), P = 0.0001 (KO-BMDN vs. WT-BMDN, N2 TAN), P = 0.0001 (KO-BMDN vs. Control, N2 TAN), P = 0.0039 (WT-BMDN vs. Control, N2 TAN). Source data are provided as a Source Data file.
