Rabbit anti cd11c

Samples of fat pads were fixed in 10% formalin solution, dehydrated, and embedded in paraffin. Serial 5‐μm sections were cut, mounted onto glass slides, deparaffinized, and rehydrated through degraded ethanol. Tissues were stained with hematoxylin‐eosin and Masson's trichrome reagents.
Immunohistochemical staining using mouse anti‐CD68 (Dako, Santa Clara, CA), rabbit anti‐CD11c (Abcam, Cambridge, UK), also known as integrin α X chain, and mouse anti‐CD206 (Abnova, Taipei, Taiwan), also known as mannose receptor C‐type 1, antibodies was performed as previously described.26, 27 Control experiments were performed by omitting the primary antibodies.

Mice in different groups were euthanized, and different tissues, including tumors, were excised and fixed in 4% paraformaldehyde. A part of tumors and DLNs were embedded, frozen, and sectioned at a thickness of 8 μm. All tissue section was treated according to the standard manufacturer’s instructions and stained with different primary antibodies: rabbit anti-CD11c (1:200), rat anti-F4/80 (1:200; Abcam), rabbit anti-CD11b (1:1000), rabbit anti–Ly-6G/Ly-6C, rabbit anti-CD4 (1:200), rabbit anti-Foxp3 (1:200), rabbit anti-CD206 (1:200), and rabbit anti–PD-L1 (1:200), followed by staining with fluorescently labeled secondary antibodies [Alexa Fluor 488–conjugated goat anti-rat IgG (H+L) (1:200; Thermo Fisher Scientific) and Alexa Fluor 633–conjugated goat anti-rabbit (H+L) (1:200; Thermo Fisher Scientific)]. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) containing mounting solution (DAPI Fluoromount-G, SouthernBiotech). All the slices were finally imaged with a confocal microscope (Leica, SP8). The other parts of tumor tissues and other organs were fixed in 4% paraformaldehyde, then embedded into paraffin, and subsequently sliced at a thickness of 5 μm. Slices were stained with hematoxylin and eosin (H&E), imaged by optical microscopy, and assessed by three independent pathologists.