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. 2020 Nov 27;48(22):12566–12576. doi: 10.1093/nar/gkaa1111

Figure 5.

Figure 5.

Metabolic labeling of RNAs in mouse gut microbiotas. (A) In-gel fluorescence scanning showing total RNA that were isolated from Acinetobacter baumannii, Bacillus subtilis, Staphylococcus aureus, Rhodococcus erythropolis, Klebsiella pneumoniae, and Salmonella typhimurium treated with 100 μM AzG for 2 h, followed by reaction with DBCO-Cy5. GelRed-stained gels demonstrate equal loading. (B) Workflow of metabolic RNA labeling of gut microbiotas with AzG in living mice. (C) In-gel fluorescence scanning showing total RNA that were isolated from mouse gut microbiotas treated with 50 μl of 20 mM AzG by oral gavage for three times with an interval of 1.5 h, followed by reaction with DBCO-Cy5. GelRed-stained gels demonstrate equal loading. (D) Confocal fluorescence microscopy images showing gut microbiotas from mice treated with 50 μl of 20 mM AzG by oral gavage for three times with an interval of 1.5 h, followed by click reaction with alkyne-Cy5. Scale bars, 2 μm. For (C) and (D), representative results are shown from three independent experiments.