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. 2022 Apr 24;113(6):1955–1967. doi: 10.1111/cas.15355

FIGURE 5.

FIGURE 5

Immunofluorescence and western blot (WB) analysis of ADP‐ribosylation factor 4 (ARF4)/Ras‐related GTP‐binding protein 10 (RAB10)/signal transducer and activator of transcription 3 (pSTAT3) on neutrophils. Blood‐derived neutrophils were treated with/without glutamate (100 μM) for 24 h. (A) Triple immunostaining of neutrophils. Green, LY6G; red, specific proteins (ARF4/RAB10/pSTAT3); blue, nucleus. Scale bar, 100 μm. (B) WB analysis of ARF4/RAB10/pSTAT3 expression on blood‐derived neutrophils treated with gradient glutamate (0, 25, and 100 μM) for 24 h. In LLC‐tumor bearing mice, neutrophils were collected from blood and tumor tissues. (C) WB analysis of ARF4/RAB10/pSTAT3 expression on neutrophils isolated from blood and tumor tissues. β‐Actin was detected as loading control. (D) Quantifications of target proteins in (B) compared to loading control. */**Differences between glutamate and control group. (E) Quantifications of target proteins in (C) compared to loading control. */**Differences between blood and tumor group. n = 3