Fig. 4: Atf3 is related to FIT regulation.

a Boxplots for the VO₂ of animals at sampling in the torpor deprivation experiment. Each dot represents one sample from one animal. Torpor-deprived animals (Dep group, n = 4) did not show an apparent change in VO₂ compared to the Mid group. b MDS plot of the TSS-based distance in the torpor-deprivation experiment. Each dot represents one sample from one animal. A clear separation between the Mid and Dep groups was not found in this analysis. c Distribution of CAGE clusters according to the mean TPM and the fold-change TPM of the Mid to Dep group. The top five up- and down-regulated torpor-deprivation-specific promoters that had annotated downstream genes are shown. d Among the torpor-specific up-regulated genes, Atf3 was the only DE gene during torpor deprivation. e The motif probability of ATF3 and TBP in torpor-specific promoters. f Estimated ΔΔCT of atf3 mRNA for each organ from normal to torpid condition. The black solid line and the dashed lines denote the median and the lower and upper 89% HPDI. The red line is drawn at zero. Heart, BAT, and the soleus muscle have lower CT during torpor, which indicates higher mRNA. See Supplementary Fig. 4b for raw CT counts. g The FIT phenotype of Atf3-KO mice (n = 8, pink lines) was compared to wildtypes (n = 4, black lines). The thick lines denote the average of either T_B or VO₂ in each group, while thin lines show individual recordings. Atf3-KO have a tendency of higher metabolism during torpor compared to wildtypes. h The minimal T_B and VO₂ during torpor were compared between Atf3-KO (pink) and wild-type mice (black). Two animals in the KO group failed to record body temperature due to equipment trouble. The three-digit numbers by the dots are the animal ID. Two KO lines, which showed higher metabolisms than wildtypes, ATF3-025, and ATF3-021, were selected for further evaluation.