Fig. 5: The molecular clock and the timing of food consumption regulate OGT and OGA protein levels. | Nature Communications

Fig. 5: The molecular clock and the timing of food consumption regulate OGT and OGA protein levels.

From: Hexosamine biosynthetic pathway and O-GlcNAc-processing enzymes regulate daily rhythms in protein O-GlcNAcylation

Fig. 5

a Western blots showing daily rhythms of OGT (top panel) and OGA protein (middle panel) in WT flies (w1118) fed ad libtum, between ZT21-3 (natural feeding time), or between ZT9-15 (unnatural feeding time). Total nuclear proteins stained by Coomassie blue (bottom panel) were used for normalization. Each biological replicate was performed independently with similar results (n = 4). b, c Quantification of daily OGT protein rhythm (n = 4; AL: p = 0.62, RF21-3: p = 3.92E−06, RF9-15: p = 0.036, RAIN; WT RF21-3 vs RF9-15: mesor p = 0.075, amplitude p = 0.25, phase p = 3.56E−06, CircaCompare) and OGA protein rhythm (n = 4; AL: p = 0.92, RF21-3: p = 0.051, RF9-15: p = 0.31; RAIN). d Western blots showing daily rhythms in OGT (top panel) and OGA protein (middle panel) in WT (w1118) and per0 flies fed between ZT21-3. Each biological replicate was performed independently with similar results (n = 4). e, f Quantification of OGT (n = 4; WT: p = 5.63E−07, per0: p = 0.68, RAIN) and OGA protein levels (n = 4; WT: p = 0.038, per0: p = 0.41, RAIN). Data are double plotted. g, h Expression of ogt mRNA in three groups of flies: WT RF21-3, per0 RF21-3, and WT RF9-15. (n = 3; WT RF21-3: p = 0.041, WT RF9-15: p = 0.0033, per0 RF21-3: p = 0.94, RAIN; WT RF21-3 vs RF9-15: mesor p = 0.84, amplitude p = 0.60, phase p = 2.36E−06, CircaCompare). i, j Expression of oga mRNA in three groups of flies: WT RF21-3, per0 RF21-3, and WT RF9-15. (n = 3; WT RF21-3: p = 0.044, WT RF9-15: p = 0.22, per0 RF21-3: p = 0.53; RAIN). The biological replicates of WT RF21-3 that are quantified in panel (h) and (j) are the same three replicates quantified in panel (g) and (i) respectively. Gray shading indicates the dark period of LD cycle. Colored bars at the top of graphs denote the feeding periods for TRF treatment groups, RF21-3 (blue) and RF9-15 (yellow). The red bars with gradient shading illustrate the representative feeding pattern for AL flies (data depicted from Fig. 2a), and the darker area indicates higher feeding activity. Data are normalized (peak=1) and are presented as mean ± SEM.

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