Figure 5. Colonization and Butyrate Rescue Diminished Oxidative Metabolism.

(A) Mitochondrial respiration indicated by MitoTracker Red CM-H₂XRos (red fluorescence; top panel) and MitoTracker Green FM (green fluorescence; middle panel) in colonocytes from CONV-R, CONV-D, B. fibrisolvens, GF, GF with 10 mM butyrate, and GF with both 10 mM butyrate and 500 µM etomoxir. (B) Quantification of mitochondrial respiration in different experimental groups. (C–D) Western blot analysis of phospho-p27 (C) and LC3-I and -II (D) with and without 10 mM butyrate in CONV-R and GF with β-actin as loading controls. (E) Western blot analysis of LC3-I and –II of GF, GF with 10 mM butyrate, and GF with 10 mM butyrate and 500 µM etomoxir. For panels A and B, a total of 3 mice per condition were used, and results are displayed as mean ± SE, with significant differences indicated (**p <0.01). For panels C–E, western blots are representative of 4 experiments using 2 CONV-R and 2 GF mice with 2 technical replicates each.