Fig 1: Gene analysis on patient-derived tumor cells highlighted high TNFα expression. A Representative immunohistochemistry images showing a CLM patient with higher S-TAMs (left) and higher LTAMs (right) in the peritumoral area. Scale bar: 30 µm. B TNFα and TGFβ expression in primary cells from patients with higher S-TAMs (n = 3) and with higher L-TAMs (n = 3). C LSD1 expression in primary cells from patients with higher S-TAMs (n = 3) and with higher L-TAMs (n = 3). Gene expressions were normalized using GAPDH as a housekeeping gene. Data are represented as mean ± SD of the average of three biological replicates (Mann–Whitney test; *p < 0.05)
Fig 2: LSD1 and riboflavin receptors are increased in M2 macrophages under inflammatory stimuli. A LSD1 expression in monocytes differentiates into M2 macrophages exposed to TNFα (20 ng/ml) for 18 h. B LSD1 expression in monocytes differentiates into M2 macrophages exposed to TGFβ (10 ng/ml) for 18 h. C Riboflavin receptors (SLC52A2 and SLC52A3) expressions in M2 macrophages exposed to TNFα (20 ng/ml) or TGFβ (10 ng/ml) for 18 h. D Riboflavin receptors (SLC52A2 and SLC52A3) expressions in M1, M2, and M2 exposed to TNFα (20 ng/ml) or TGFβ (10 ng/ml) for 18 h. Gene expressions were normalized using GAPDH as a housekeeping gene. Data are represented as mean ± SD of the average of three biological replicates. **p < 0.01; ***p < 0.001 Mann–Whitney test when comparing two groups. *p < 0.05; **p < 0.01 One-way ANOVA when comparing three groups
Fig 3: Role of riboflavin in driving TAMs morphology and polarization. A Schematic overview of the biochemical link between Riboflavin (vitamin B2) and LSD1 enzyme. B At left, riboflavin levels expressed as peak area in S- and L-TAM populations in the 14 TAM pairs. At right, LSD1 activity of S- and L-TAM pairs obtained from 8 CLM-patients expressed as OD. C LSD1 expression analyses in S- and L-TAMs (n = 4 CLM patients). Gene expressions were normalized using GAPDH as a housekeeping gene. D At left, LSD1 expression levels in M1 and M2 in-vitro polarized monocytes. At right, LSD1 activity of M1 and M2 polarized monocytes expressed as OD. Gene expressions were normalized using GAPDH as a housekeeping gene. Gene expression data are represented as mean ± SD of the average of three biological replicates. *p < 0.05, **p < 0.01 Wilcoxon Mann–Whitney test
Supplier Page from Abcam for KDM1/LSD1 Activity Quantification Assay Kit (Colorimetric)