Skip to main content
. 2022 Apr 18;9(17):2200435. doi: 10.1002/advs.202200435

Figure 3.

Figure 3

Internalization, anti‐ROS, anti‐inflammation, and anti‐polarization effects and transcriptome analysis in RAW264.7 cells. a) Scheme of the experimental procedures to evaluate oxidative stress, inflammation, and polarization in RAW264.7 cells. b) Representative fluorescence images of time‐dependent cellular uptake behaviors of MTem‐Rho (Rho concentration: 10 mm equivalent to those of free Rho) or 0.5, 1, and 2 h into RAW264.7 cells. Green: Phalloidin; Blue: DAPI; Red: MTem‐Rho signal. The scale bars are 25 µm. c) Flow cytometric curves of internalization of MTem‐Rho. d) Quantitative analysis of fluorescent intensity according to the flow cytometry. Data are presented as mean ± SD; n = 3. **p < 0.01. e) Representative ROS images of RAW264.7 cells by DCFH‐DA staining after different treatments. The scale bar is 50 µm. f) Flow cytometer diagram of DCF of different treatments. g) Quantitative analysis of the average fluorescent intensity according to the flow cytometer. RNA levels of h) IL‐1β and i) MMP‐9 in LPS‐induced RAW264.7 cells treated with different treatments after 24 h, analyzed by Q‐PCR. j) Western Blotting images of p38 MAPK, p‐p38 MAPK, GAPDH protein expressions (LPS induced for 1 h), NF‐κB p‐p65, MMP‐9, IL‐1β, and GAPDH protein expressions (LPS induced for 24 h) in RAW264.7 cells. k) CD80 and CD206 flow cytometry curves of LPS‐induced RAW264.7 cells. l) Corresponding flow cytometric analysis of CD80 and CD206 in RAW264.7 cells. Data were presented as mean ± SD; n = 3. ns: not significant, p > 0.05, ##p < 0.01 versus the Normal group, and **p < 0.01 versus the LPS group. m) Venn diagram demonstrating the intersection set of DEAS and DEG. n) Functional analyses from MTem/Los‐associated DEAS events in RAW264.7 cells, including GO and KEGG. o) Heat maps of significantly down‐regulated genes in the RAW264.7 cells after MTem/Los treatment (fold change ≥ 1.5 and p < 0.05).