Fig 2: NK cell depletion reverses gMoxi-mediated tumour regression. Lewis lung cancer (LLC) cells were inoculated on the upper back of C57BL6 mice on day 0, and gMoxi was performed at the acupoint Zusanli (ST36) with three moxa cones every other day from day 1. A total of 100 µg of PK136 per mouse was administered by IP injection on days 0, 7 and 14. Mice were killed on day 21, and tumours and spleens were isolated and analysed by flow cytometry. A, The proportion of CD3-NKp46+ NK cells in spleens; B, the proportion of CD3+CD4+ T cells in spleens; C, the proportion of CD3+CD8+ T cells in spleens; D, the proportion of CD3-NKp46+ NK cells in tumours; E, the proportion of CD3+CD4+ T cells in tumours; F, the proportion of CD3+CD8+ T cells in tumours; G, tumour growth curve; H, tumour photograph; I, tumour weight; J, splenocytes were co-cultured with YAC1 cells at 10:1, and tumour lysis was detected by apoptosis analysis. Independent experiments were repeated twice. *P < .05; **P < .01; ***P < .001; NS, non-statistically significant

Fig 3: Differences in the cellular metabolic status and mitochondrial activity of splenic NK cells between DHA-fed and control mice at day 7 post-MCMV infection. A–E Flow cytometric analysis and cumulative results depicting the gMFI of CD71 (A) and CD98 (B) and the uptake of 2-NBDG (C), MitoTracker (D) and tetramethylrhodamine, methyl ester (TMRM) (E) by splenic NK cells (CD3-CD19-NK1.1+NKp46+) from control versus DHA-fed mice at day 7 postinfection. For each experiment, n = 7 pooled from 2 independent experiments. Each symbol represents an individual mouse, and the blue dots and red squares represent control and DHA-enriched diet-fed mice, respectively. Error bars represent interquartile ranges; *p < 0.05; **p < 0.01; ***p < 0.001

Fig 4: RocA increases NK cell infiltration in NSCLC. LLC cells were subcutaneously inoculated onto the upper back of C57BL6 mice on day 0, and 1 mg/kg of RocA was administered by i.p. injection every 2 days from day 3. Mice were sacrificed on day 21, and tumors were excised. The proportion of NK cells in tumors were analyzed by flow-cytometry (A-B). Tumor tissues were stained with anti-NKp46 antibody and DAPI (C), and the NKp46+ NK cells were quantified in tumor tissues (D). The proportion of NK cells in spleens in LLC tumor-bearing mice (E) and normal mice without LLC tumor (F) were analyzed by flow-cytometry.

Fig 5: Mechanism of action of IRS661. (A) Schematic representation of the experimental setup. Mice were infected intranasally with 1,000 TCID50 of influenza virus X31 and treated with IRS661 or PBS intranasally (15 nmol in 30 µl) on day 4 after infection. Usually, BALF was isolated on day 6 after infection. Colored dots mark days when mice were treated with aPDCA-1 (200–500 µg), 1A8 (150–300 µg), aSiglec-H (200–500 µg), or aCCR2 (20 µg) depletion antibody i.p. (B) WT 129S7 mice were infected with X31 and treated with IRS661 or PBS intranasally (15 nmol in 30 µl), i.p. (50 nmol in 100 µl), or intravenously (50 nmol in 100 µl) 4 d later. IFN-a concentrations were quantified by ELISA in BALF taken on day 5 after infection. n = 3–11 mice per group. (C) Flow cytometry plots representing the gating strategy for cell analysis in Fig. 2 B. Lineage channel includes CD3, CD19, and NKp46. FSC, forward scatter; SSC, side scatter. (D) WT 129S7 mice were infected with 1,000 TCID50 X31 and treated with IRS661, control ODN 2, or PBS intranasally. Survival was monitored for 14 d. n = 17–18 mice per group. (E and F) Viral loads in lungs of X31-infected mice treated with IRS661 or PBS (E; n = 3–7 mice per group) and/or aPDCA-1 antibody (F; n = 6–7 mice per group) were determined by RT-qPCR or TCID50 assay at days 2, 3, 5, and 7 (E) or day 6 (F) after infection. (G) WT C57BL/6 mice were infected with 10,000 TCID50 X31 and treated with IRS661 or PBS intranasally. Survival was monitored for 14 d. n = 23 mice per group. (H) WT C57BL/6 mice were infected with 5 TCID50 PR8 and treated with IRS661 or PBS intranasally. Survival was monitored for 14 d. n = 13–14 mice per group. (I) WT Balb/c mice were infected with 10,000 TCID50 X31 and treated with IRS661 or PBS intranasally. Survival was monitored for 14 d. n = 16 mice per group. (J) WT 129S7 mice were infected and treated with IRS661 or PBS and aCCR2 antibody or isotype control as depicted in A. Survival was monitored for 14 d. n = 18–20 mice per group. (K) WT 129S7 mice were infected with X31 and treated with IRS661 or PBS. BALF and lung were isolated on day 8 after infection and checked for CD44+ Pentamer+ CD8+ T cells by flow cytometry. n = 8–10 mice per group. (L) Rag-2 KO (129) mice were infected with 750 TCID50 X31 and treated with IRS661 or PBS intranasally. Survival was monitored for 14 d. n = 10–11 mice per group. Data are pooled from at least two independent experiments, and each symbol in the bar charts represents a biological replicate. Statistical analysis was performed using unpaired t test (B and K), log-rank test (D, G–J, and L), 2-way ANOVA with Tukey’s multiple comparisons test (E), or ordinary one-way ANOVA with Tukey’s multiple comparisons test (F). **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.