Fig. 5: Defect of TYMS rescues the pro-inflammatory phenotype of RRP1 -deficient macrophages.
From: The RNA-binding protein RRP1 brakes macrophage one-carbon metabolism to suppress autoinflammation
RT-qPCR detection of Il-1b (a) and Il6 (b) mRNA levels in RRP1 KO RAW 264.7 cells upon TYMS silencing and IL-1β stimulation for the indicated hours. c Western blot detection of RRP1 and TYMS protein levels in RRP1 KO RAW 264.7 cells upon TYMS silencing targeted by two distinct siRNAs. GAPDH serves as loading control. d RT-qPCR detection of IL-1B mRNA levels in negative control (NC)- and RRP1, TYMS or double-silenced THP-1 cells stimulated with IL-1β for the indicated hours. e ELISA detection of IL-1β levels in the supernatant of NC- and RRP1, TYMS or double-silenced THP-1 cells stimulated with IL-1β for the indicated hours. f Heatmap of 1C-metabolites in RRP1 KO RAW 264.7 cells upon TYMS silencing and IL-1β stimulation for the indicated hours. Color indicates normalized intensities (z-score). g ELISA detection of SAM, methionine (Met), THF levels in RRP1 KO RAW 264.7 cells upon TYMS silencing and IL-1β stimulation for the indicated hours. h RT-qPCR detection of Il-1b and Il6 mRNA levels in RRP1 KO RAW 264.7 cells pre-treated with Raltitrexed in the indicated concentrations and IL-1β stimulation for the indicated hours. i Western blotting for TMYS in RRP1-silenced BMDMs stimulated by IL-1β for the indicated hours. GAPDH serves as loading control. Western blotting for TMYS in the WT and RRP1 KO RAW 264.7 cells upon poly(I:C) (j) or murine IL-6 (k) stimulation for the indicated hours. GAPDH serves as loading control. Western blotting data are representative of three independent experiments (c, i, j, k). Metabolome data are from three biologically independent samples. RT-qPCR data are presented as means ± SD (n = 3) of biologically samples from three independent experiments with student’s t test (two-tailed unpaired) (a, b, d, e, g, h). Source data are provided as a Source Data file.
