Effect of functionalized nanocellulose on macrophage polarization
Effect of functionalized nanocellulose on macrophage polarization
The transcriptome analysis was carried out to evaluate the differentially expressed genes (DEGs) in various experimental groups after 24 hours of treatment. Total RNA extraction from murine macrophage/monocytes (RAW 264.7 cells) The RAW 264.7 cells were cultured in DMEM, supplemented with 10% FBS and 1% antibiotics in a humidified atmosphere. The control cells recieved PBS, followed by LPS (100 ng/mL), IL-4 (100 ng/mL), CN (100 ug/mL), and CND (50 ug/mL) Excel-based Differentially Expressed Gene Analysis (ExDEGA) software tool provided by eBiogen Corp., Republic of Korea. The DEGs expression was first filtered based on gene category settings. This was followed by the building of multiple comparisons, grouping, and analyzing the scattered/volcano plotting with fold change value of each DEGs (Log2 fold change, p < 0.05). Next, we performed the clustering analysis, GSEA, and STRING analysis to understand the interactions between most significant genes responsible for macrophage polarization among various groups Total RNA isolation using Trizol method and the RNA purity was quantified using Nanodrop spectrophotometer. Briefly, the RAW 264.7 cells were growth for 24 h, followed by treatment with various compositions up to 24 hours. After 24 hours, the cells were washed with 1x PBS and the total RNA (1 mL of Trizol /6-well/ 200000 cells) was extracted  Poly(A) RNA selection from total RNA, followed by magnetic beads-based separation which ensures highly specific poly(A) enrichment