Equal amounts of protein lysates (20 µg) extracted from cells transfected with NC and small interfering (si) IL4I1 were subjected to SDS-PAGE and transferred to PVDF membranes. Following incubation with 5% skimmed milk for 1 h at room temperature (RT), PVDF membranes were exposed to primary antibodies targeting IL4I1 (1:1,000, ab222102, Abcam), CD11B (1:1,000, ab133357, Abcam), CD204 (1:1,000, ab271070, Abcam), CD206 (1:2000, 60143-1-Ig, Proteintech), CD163 (1:1,000, ab182422, Abcam), CD86 (1:1,000, ab239075, Abcam), and beta-tubulin (1:2000, DF7967, Affinity) overnight. Subsequently, PVDF membranes were incubated with Goat Anti-Rabbit/Mouse IgG (H+L) HRP (1:5,000, Affinity) for 1 h. Chemical imaging was developed using ECL solutions (Epizyme, Shanghai, China). Pictures were obtained using a Bio-Rad imaging system.
The extraction of total RNA from cells transfected with NC and siIL4I1 was done through an RNA Purification Kit (B0004D, EZBioscience, USA). Subsequently, reverse transcription was carried out using RNA (1 μg). After ward, RT-qPCR was done using the SYBR qPCR Master Mix kit (Epizyme, Shanghai). Amplification was detected using QuantStudio (Thermo Scientific, USA). The primer sequences we used are listed inTable 3
The extraction of total RNA from cells transfected with NC and siIL4I1 was done through an RNA Purification Kit (B0004D, EZBioscience, USA). Subsequently, reverse transcription was carried out using RNA (1 μg). After ward, RT-qPCR was done using the SYBR qPCR Master Mix kit (Epizyme, Shanghai). Amplification was detected using QuantStudio (Thermo Scientific, USA). The primer sequences we used are listed in