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. 2022 Feb 1;132(3):e144828. doi: 10.1172/JCI144828

Figure 4. Role of TAS2R43 in the effect on E. coli growth of CSNs from patients with obesity coli growth of CSNs from patients with obesity stimulated with bitter compounds.

Figure 4

(A) E. coli CFU following treatment with the supernatant of obese patients’ jejunal crypts (n = 5–14) stimulated for 30 minutes with bitter compounds (compound-CSN) or Krebs (Krebs-CSN). Crypt-independent effects of bitter agonists are shown as well (bacteriostatic control). All results are expressed as the percentage of Krebs-CSN with or without vehicle. Compounds were grouped by their TAS2R activation profile: generalists (>5 TAS2Rs); intermediates (3 or 4 TAS2Rs), and specialists (1 or 2 TAS2Rs). Test concentrations and abbreviations are given in Table 1. (B) Western blots showing expression of α-defensin 5 or REG3A in lysates from crypts from obese individuals. Crypts were treated for 30 minutes with bitter agonists or Krebs. Vertical line indicates that the lanes were run on the same gel but were noncontiguous. Summary of Western blot analyses for the effect on (C) α-defensin 5 or (D) REG3A protein expression (n = 6–8). (E and F) Effect of TAS2R43 deletion/amino acid polymorphisms. (E) Effect on E. coli growth of the supernatant of crypts from TAS2R43+ and TAS2R43– patients with obesity. Crypts were stimulated for 30 minutes with the TAS2R43 agonists aloin (30 μM, n = 11 TAS2R43+, n = 4 TAS2R43–); quinine (0.1 mM, n = 4 TAS2R43+, n = 2 TAS2R43–); or DB (0.1 mM, n = 4 TAS2R43+, n = 3 TAS2R43–). (F) Effect on E. coli growth of the supernatant of crypts from obese patients with a TAS2R43 genotype that is highly sensitive (n = 5 TAS2R43+ W); mildly sensitive (n = 5 TAS2R43+ S); or not sensitive (n = 4 TAS2R43–) to aloin. Data represent the mean ± SEM, and single values are plotted. Statistical significance was determined using a mixed model with patient as the random effect and, for Western blot analysis, a paired Student’s t test with Bonferroni-Holm adjustment. *P < 0.05, **P < 0.01, and ***P < 0.001, versus Krebs-CSN or the vehicle control (100% control); #P < 0.05 and ##P < 0.01, versus the bacteriostatic control.