Fig. 3: TUT7 decreases Zc3h12a mRNA stability in response to LPS challenge.

a RAW 264.7 cells were treated with 100 ng/ml LPS for 2 h and cell lysates were incubated with control IgG or protein A-agarose beads-conjugated anti-TUT7 antibody at 4 °C for 4 h. TUT7-interacting RNAs were extracted followed by RT-PCR (upper panel). Immunoprecipitation of TUT7 was examined by immunoblotting (lower panel). b–e Control (shControl) and Tut7-knockdown RAW 264.7 cells and wild-type and Tut7^−/− BMDMs were treated with 100 ng/ml LPS for different periods of time. The expression of Regnase-1 protein (b, d) and Zc3h12a mRNA (c, e) were analyzed by immunoblotting and RT-qPCR, respectively. f Control- and shTut7-expressing RAW 264.7 cells were treated with 100 ng/ml LPS for 2 h followed by treatment with 5 μg/ml of actinomycin D for the indicated periods of time. Regnase-1 mRNA expression was analyzed by RT-qPCR. g Tut7-knockdown RAW 264.7 cells were reconstituted with either hTUT7 or hTUT7(DADA) and challenged with 100 ng/ml of LPS for 2 h. Cells were collected, and the expression of the indicated proteins and mRNAs were analyzed by immunoblotting and RT-qPCR, respectively. h Control- and shZc3h12a-expressing Tut7^−/− BMDMs were stimulated with 100 ng/ml LPS for the indicated periods of time. Total RNAs were prepared and the mRNA expression of the indicated genes was measured by RT-qPCR. Data presented are representative of three independent experiments with triplicates in each experiment (error bars, mean ± S.D.). The p values were obtained from two-tailed Student’s t test and are shown in the figure if p < 0.05. Source data are provided as a Source Data file.