Journal: Nature communications

Article Title: Refractoriness of STING therapy is relieved by AKT inhibitor through effective vascular disruption in tumour.

doi: 10.1038/s41467-021-24603-w

Figure Lengend Snippet: Fig. 6 cGAMP fails to induce tumour EC apoptosis and anti-tumour growth in MMTV-PyMT spontaneous breast cancer. a–c Diagram depicting treatment and sampling in 9 weeks-old MMTV-PyMT mice. Representative images and comparisons of apoptosis in tumour ECs and whole tumour cells (whole cells). Scale bars, 1.0 mm (yellow bars) and 100 μm (white bars). n = 5 mice/group from two independent experiments. Vertical bars indicate mean ± SD. d–f Diagram depicting treatment and sampling in 9 weeks-old MMTV-PyMT mice. Comparison of tumour growths. n = 7 mice/group from two independent experiments. Plots and bars indicate mean ± SD. Plot indicates each individual tumour growth. g Diagram depicting generation of an orthotopic implanted breast tumour in syngeneic FVB mice. h-j Diagram depicting treatment and sampling in implanted breast tumour mice. Representative images and comparisons of apoptosis in tumour ECs (white arrowheads) and whole tumour cells. Scale bars, 1.0 mm (yellow bars) and 100 μm (white bars). n = 5 mice/group from two independent experiments. Vertical bars indicate mean ± SD. k–m Diagram depicting treatment and tumour growth in implanted breast tumour mice. Comparisons of tumour growths. n = 7 mice/group from two independent experiments. Plot and bars indicate mean ± SD. Plot indicates each individual tumour growth. n Comparison of reduction of tumour volume by cGAMP treatment between spontaneous and implanted breast tumours at day 47 after the implantation. o, p Diagram depicting treatment and sampling 6 h later in 9-week-old MMTV-PyMT mice and its implanted breast tumour mice 10 day (d10) after implantation. Comparisons of TNFα and IFNγ levels in tumour lysates. n = 5 mice/group from four independent experiments. Horizontal bars indicate mean ± SD. P values by two-tailed t-test (c, e, j, l, n) or Welch’s one-way ANOVA test followed by Dunnett’s T3 test (p). ns, not significant. Source data are provided as a Source Data file.

Article Snippet: For blocking TNFα, anti-TNFα monoclonal antibody (clone XT3.11, 15 mg/kg of body weight, diluted in 100 μl PBS; #BE0058, BioXcell) or isotype control antibody (clone HRPN, 15 mg/kg of body weight diluted in 100 μl PBS, Rat IgG1; #BE0088, BioXcell) was intraperitoneally (i.p.) administered. i.t. injection of TNFα (210 ng/ 70 μl of PBS; Sigma), IFNγ (350 ng/70 μl of PBS; R&D systems) or the combination (in 70 μl of PBS) was performed at day 13 after tumour implantation.

Techniques: Sampling, Comparison, Two Tailed Test

Journal: Nature communications

Article Title: Refractoriness of STING therapy is relieved by AKT inhibitor through effective vascular disruption in tumour.

doi: 10.1038/s41467-021-24603-w

Figure Lengend Snippet: Fig. 9 AKT inhibitor sensitizes cultured HUVECs to TNFα-induced apoptosis. a Immunoblot analysis of FOXO1, phosphorylated FOXO1 at threonine 24 [pFOXO1 (T24)], AKT, phosphorylated AKT at serine 473 [pAKT (S473)] and α-tubulin in cultured HUVECs at 30 min after indicated treatments. Three independent experiments showed similar findings. b Representative images of FOXO1 subcellular localization in the ECs of LLC tumour. PBS (70 μl), cGAMP (14 μg/70 μl) or AKTi (50 mg/kg) was injected 3 h before sampling. White arrowheads indicate nuclear localization of FOXO1. Scale bars, 50 μm. Five independent experiments showed similar findings. c, d Representative images and comparison of apoptosis in cultured HUVECs treated with indicated agents. TNFα (200 ng/ml), IFNγ (300 ng/ml) or AKTi (10 μM) was added and incubated for 6 h. Scale bars, 100 μm. Dots indicate values from four independent experiments. Vertical bars indicate mean ± SD. P values by Welch’s one-way ANOVA test followed by Dunnett’s T3 test. ns, not significant. e–g Viabilities of cultured HUVECs, LLC cells, and MMTV-PyMT tumour cells by indicated treatments. TNFα (200 ng/ml), IFNγ (300 ng/ml) or AKTi (10 μM) was added and incubated for 24 h. Dots indicate values from six independent experiments. Vertical bars indicate mean ± SD. P values by Welch’s one- way ANOVA test followed by Dunnett’s T3 test. ns, not significant. Source data are provided as a Source Data file.

Article Snippet: For blocking TNFα, anti-TNFα monoclonal antibody (clone XT3.11, 15 mg/kg of body weight, diluted in 100 μl PBS; #BE0058, BioXcell) or isotype control antibody (clone HRPN, 15 mg/kg of body weight diluted in 100 μl PBS, Rat IgG1; #BE0088, BioXcell) was intraperitoneally (i.p.) administered. i.t. injection of TNFα (210 ng/ 70 μl of PBS; Sigma), IFNγ (350 ng/70 μl of PBS; R&D systems) or the combination (in 70 μl of PBS) was performed at day 13 after tumour implantation.

Techniques: Cell Culture, Western Blot, Injection, Sampling, Comparison, Incubation