Journal: Life Science Alliance
Article Title: Reprogramming of breast tumor–associated macrophages with modulation of arginine metabolism
doi: 10.26508/lsa.202302339
Figure Lengend Snippet: (A) Levels of BH 4 produced by THP-1–derived M0-, M1-, and M2-TAMs after being treated with vehicle (DMSO) or SEP (100 μM) for 3 d (n = 6). BH 4 levels were measured with ELISA and normalized against the total protein levels. One-way ANOVA with a post hoc test (Tukey’s test) was performed to measure the significance of the mean difference between treatment groups. (A, B) Phalloidin staining and SEM imaging of THP-1–derived M0-, M1-, and M2-TAMs treated as in (A) (n = 3). SEM images are shown at different magnifications. (A, C) Western blot analysis of TAM subsets treated with a vehicle or SEP as in (A), and β–actin was used as the internal loading control (n = 5). (D) Quantification of the Western blot results based on the expression of TLR2 (M1 marker) versus CD206 (M2 marker) normalized against β–actin signal and presented as fold differences. (E) Immunofluorescence imaging of THP-1–derived TAMs after treatments as shown above, and stained for an M1 marker (green, TNFα) versus M2 marker (red, CD206) and counterstained with DAPI (blue) (n = 3). (F) Levels of secreted cytokines, type 1: TNFα (top left), IL1β (top right), and IL6 (bottom left) versus type 2: TGFβ (bottom right), for M1- versus M2-TAMs treated with a vehicle versus SEP (n = 6) measured with ELISA. Error bars: ±SEM. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001; and ns, P > 0.05.
Article Snippet: The ELISA kits used were as follows: IL12 (Cat. No. D1200; R&D Systems), IL6 (Cat. No. D6050; R&D System), IL1β (Cat. No. DLB50; R&D System), TNFα (Cat. No. DTA00D; R&D System), IL10 (Cat. No. D1000B; R&D Systems), and TGFβ (Cat. No. ab108912; Abcam).
Techniques: Produced, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining, Imaging, Western Blot, Control, Expressing, Marker, Immunofluorescence