Journal: Frontiers in immunology

Article Title: YTHDF1 Negatively Regulates Treponema pallidum -Induced Inflammation in THP-1 Macrophages by Promoting SOCS3 Translation in an m6A-Dependent Manner.

doi: 10.3389/fimmu.2022.857727

Figure Lengend Snippet: FIGURE 1 | m6A methylation: the m6A writer METTL3 and the reader YTHDF1 are upregulated in TP-infected macrophages. (A) Colorimetric quantification of m6A methylation in RNA from THP-1 cells with or without TP infection. (B, C) Western blot analysis of METTL3, METTL14, and YTHDF1 in THP-1 cells with or without TP infection. GAPDH was used as a control. *P < 0.05, **P < 0.01. (D, E) Immunofluorescence analysis of YTHDF1 or METTL3 in CD68+ macrophages in secondary syphilitic lesions and paired non-lesional skin tissue. Scale bar: 20 mm or 5 mm.

Article Snippet: For immunofluorescence analysis of the tissues, the sections were permeabilized with 0.25% Triton X-100 for 0.5 h and blocked with 5% goat serum for 1 h. Tissue sections were then incubated with primary antibodies against CD68 (Cat. No. 66231-2-Ig, Proteintech, Wuhan, Hubei, China) and YTHDF1 (Cat. No. 17479-1-AP, Proteintech) at 4°C overnight, followed by April 2022 | Volume 13 | Article 857727 incubation with the secondary antibody, Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) (A21206, Life Technologies) or Alexa Fluor® 594 donkey anti-mouse IgG (H+L) (A21203, Life Technologies).

Techniques: Methylation, Infection, Western Blot, Control