Ultra leaf purified anti mouse ly 6g antibodies

After the 4T1 cancer cell mouse model was treated with CXCL2 plasmid DNA (pDNA) in combination with HVJ-E treatment, HVJ-E treatment, CXCL2 pDNA treatment, or PBS treatment for 3 weeks, lung sections were fixed with 4% paraformaldehyde solution and blocked with 5% BSA. The sections were stained with Ultra-LEAF Purified anti-mouse Ly-6G antibodies (1A8, 127649, Biolegend). The secondary antibodies included an Alexa Fluor 488-conjugated rabbit anti-rat IgG (Life Technologies, Carlsbad, CA, USA). The sections were mounted in Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA) and imaged with a confocal laser scanning microscope (LSM880 with Airyscan, Zeiss, Jena, Germany) equipped with the ZEN software program.

For the neutrophil-blocking experiment, 4T1 tumor-bearing mice were pretreated with an intraperitoneal injection of Ultra-LEAF Purified anti-mouse Ly-6G antibodies (100 μg in 50 μL of PBS, 1A8, 127649, Biolegend) or IgG from rat serum (100 μg in 50 μL of PBS, 14131, Sigma-Aldrich, Japan) six times 24 h before being IT injected with CXCL2 plasmid DNA (200 μg in 50 μL of PBS), HVJ-E (1,000 HAU in 50 μL of PBS), CXCL2 in combination with HVJ-E (200 μg and 1,000 HAU in 50 μL of PBS), or PBS (50 μL) once, followed by five additional treatments of HVJ-E (1,000 HAU in 50 μL of PBS) or PBS (50 μL) treatment. After the final injection, the tumor size was measured every 2 days.

A total of 1 × 10⁶ viable 4T1 breast cancer cells (in 50 μL of PBS) were intradermally injected into the right fourth mammary gland of BALB/c mice. Tumor-bearing mice were pretreated with an intraperitoneal injection of Ultra-LEAF Purified anti-mouse Ly-6G antibodies (100 μg in 50 μL of PBS, 1A8, 127649, Biolegend), IgG from rat serum (100 μg in 50 μL of PBS, 14131, Sigma-Aldrich, Japan), or PBS (50 μL) 24 h before tumor treatment. Then, the mice were treated with an intraperitoneal injection of InVivoPlus anti-mouse PD-1 (CD279) (250 μg in 50 μL of PBS, RMP1-14, BP0146, BioXcell), control IgG from rat serum (250 μg in 50 μL of PBS, 14131, Sigma-Aldrich, Japan), or PBS (50 μL) and one IT injection of C/H, followed by HVJ-E (200 μg and 1,000 HAU in 50 μL of PBS) or PBS (50 μL), followed by treatments of HVJ-E (1,000 HAU in 50 μL of PBS) or PBS (50 μL) treatment. The tumor volume was measured every 2 days.