Figure 7.

Kaempferol functions as a potent natural TRPC5 inhibitor. (A) Chemical structures of ML-204 and kaempferol. (B) Averaged Ca²⁺ imaging time courses (F340/380) with representative calcium response traces in MKN-45 and DLD-1 cells pretreated with or without kaempferol, following stimulation with 50 μM Gd³⁺. (C, D) Immunoblot analysis of AKT, p-AKT, PI3K, and mTOR expression in MKN-45 and DLD-1 cells treated with varying concentrations of kaempferol. Data are expressed as means ± SD, *P < 0.05, **P < 0.01, ***P < 0.001 (vs. DMSO group). (E, F) Immunoblot analysis of TRPC5 protein levels in MKN-45 and DLD-1 cells treated with different concentrations of kaempferol. Data are expressed as means ± SD. (G, H) DARTS detection via immunoblot in MKN-45 and DLD-1 cells. Jurkat cell lysates were incubated with kaempferol (200 μM) or DMSO, followed by digestion with Pronase at protein ratios of 1:10000, 1:3000, 1:1000, 1:300, and 1:100. Data are expressed as means ± SD, *P < 0.05, **P < 0.01, ***P < 0.001 (vs. DMSO group). (I, J) CETSA detection via immunoblot in MKN-45 and DLD-1 cells. Jurkat cell lysates were incubated with kaempferol (200 μM and 400 μM) or DMSO, followed by a 3-minute incubation at a temperature gradient of 42-64°C.