Pe anti mouse cd163

Tumors were collected from tumor‐bearing nude mice and then were digested by Trypsin (Solarbio), Collagenase (Yeasen), Hyaluronidase (Yeasen), and DNase I (Beyotime). After digestion, the suspension was filtered through a 70 µm strainer to prepare the single‐cell suspension. Cells were stained with APC‐Cy7 Anti‐Mouse CD45 (BD Biosciences), PerCP/Cyanine5.5 antimouse/human CD11b (Biolegend), APC antimouse CD68 (Biolegend), and PE antimouse CD163 (Biolegend) for 30 min at 4 °C, protect from light. Cells were then washed and analyzed in a BD FACSCelesta Multicolor Flow Cytometer (BD Biosciences). Data were analyzed by FlowJo Software 10.8.1.

The HES used in this study was obtained from Tokyo Chemical Industry (Tokyo, Japan), dissolved in distilled water, and administered orally at a dose of 5 or 10 mg/kg/day daily to mice.
For the flow cytometry analysis, FITC anti-mouse CD11b, AF647 anti-mouse F4/80, PE anti-mouse CD163, PerCP/Cyanine5.5 anti-mouse CD206, FITC anti-mouse CD45, and PE anti-mouse CD86 antibodies were purchased from BioLegend (San Diego, CA, USA). Specific antibodies against phospho-forkhead box O3a (p-FoxO3a), FoxO3a, phospho-AKT (p-AKT), AKT, phospho-mTOR (p-mTOR), mTOR, phospho-p70S6 kinase (p-p70S6K), and p70S6K were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Fbx32, MuRF1, myostatin, and MyoD were purchased from Abcam (Cambridge, UK). Antibodies against phosphoinositide 3-kinase (PI3K), myogenin, and myocyte enhancer factor 2 (MEF-2) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

For fluorescence-activated cell sorting analysis, 10⁵ peritoneal macrophages or THP-1-induced M0 macrophages (CTR group and UVB group) were isolated and resuspended in 100 µl PBS, and antibodies were added to the corresponding cells. Flow cytometric analysis was performed by FACSCanto II (BD Biosciences) using Flow Jo software. The following antibodies were used: PE/Cyanine7 anti-human CD206 (BioLegend, 321,124), FITC anti-human CD86 (BioLegend, 374,204), APC anti-human CD163 (BioLegend, 333,609), PE/Cyanine7 anti-mouse CD86 (BioLegend, 105,014), FITC anti-mouse CD206 (BioLegend, 141,704), PE/Cyanine7 anti-mouse CD204 (Invitrogen, 2,263,083), and PE anti-mouse CD163 (BioLegend, 156,703).