Fig 2: Complex spikes and complex spike - simple spike interaction are affected by TRPC3 mutations.(A) Representative PC recording traces and complex spikes shape of Z– (light black) and Z+ (dark black) PCs in the control mice. (B) Top half, comparison of complex spike firing rates in TRPC3Mwk/- (red) and pcp2Cre;TRPC3fl/fl (green) mice versus their respective littermate controls for Z– PCs (TRPC3Mwk/-: t68=2.68, p=0.009; pcp2Cre;TRPC3fl/fl: t54=2.50, p=0.016) and Z+ PCs (TRPC3Mwk/-: t42=1.56, p=0.126; pcp2Cre;TRPC3fl/fl: t54=1.41, p=0.164). Bottom half, comparison of complex spike firing rates in pcp2Cre;TRPC3fl/fl;EAAT4GFP/- (blue) and pcp2CreERT2;TRPC3fl/fl (orange) mice versus their respective controls for Z– PCs (pcp2Cre;TRPC3fl/fl;EAAT4GFP/-: t28=3.49, p=0.002; pcp2CreERT2;TRPC3fl/fl: t53=-0.940, p=0.352) and Z+ PCs (pcp2Cre;TRPC3fl/fl;EAAT4GFP/-: t20=3.03, p=0.007; pcp2CreERT2;TRPC3fl/fl: t44=0.448, p=0.656). (C) Raster plots of simple spike activity around the occurrence of each complex spike (−100 to +300 ms). These peri-complex splike time histograms can, based on post-complex spike activity, be divided into one of four types: normal (no change), facilitation, suppression and oscillation. (D) The distribution of post-complex spike response types for Z– and Z+ PCs, in TRPC3Mwk/-, pcp2Cre;TRPC3fl/fl, pcp2Cre;TRPC3fl/fl;EAAT4GFP/- and pcp2CreERT2;TRPC3fl/fl mice. Data are represented as mean ± s.d., for values see Source data, * means p<0.05 and **p<0.01.10.7554/eLife.45590.018Figure 5—source data 1.Source data for Figure 5 and supplement.Source data for the in vivo recordings in TRPC3Mwk/-, pcp2Cre;TRPC3fl/fl, pcp2Cre;TRPC3fl/fl;EAAT4GFP/- and pcp2CreERT2;TRPC3fl/fl mice, complex spikes data.

Fig 3: The decrease of both TRPC4 and TRPC5 in the hippocampi of Cx3cr1CreER IL-10−/−mice. a RT-PCR results of TRPC isoforms in the hippocampus (n = 3 in each group, t-test). The marked significance is the comparison of the same isoform in the Cx3cr1CreER IL-10−/− with the littermate. b western blotting bands of TRPC1, TRPC3, TRPC4, and TRPC5 in littermates and in Cx3cr1CreER IL-10−/−mice. Bar graphs represent densitometric plots of protein expression in littermate and Cx3cr1CreER IL-10−/−mice in TRPC1 (c), TRPC3 (d), TRPC4 (e), and TRPC5 (f). Each dot represents a mouse. Bars represent mean ± SEM. n = 4 in each group, Significant differences were established by t-test. g Expression of TRPC1, TRPC3, TRPC4, and TRPC5 in the CA3 region of mouse hippocampal slices. Immunofluorescence images were captured with a 20 × objective. Green, immunoreactivity of TRPC1, TRPC3, TRPC4, and TRPC5; blue, nuclei stained with DAPI. Merged images of each TRPC isoform and DAPI staining. *P < 0.05, **P < 0.01, ***P < 0.001

Fig 4: The interaction between TRPC3 and SIRT1. A, Co‐localization of TRPC3 and SIRT1. Cells are co‐stained with antibodies against TRPC3 (Alexa Fluor 488, green) and SIRT1 (Alexa Flour 564, red). Arrow indicates co‐localization. B, Interaction of TRPC3 with SIRT1 in mammalian cells. C, Endogenous TRPC3 binds to SIRT1 in MCFs. D, The interaction of endogenous TRPC3 with SIRT1 in MCFs. E, Schematic illustration of the domains of TRPC3 and its association with SIRT1. F, The C‐terminal tail (659‐836) of TRPC3 interacts with SIRT1. G, H, Analyse of the binding affinity between full‐length SIRT1 and TRPC3‐C (G) or TRPC3‐N (H) by bio‐layer interferometry (BLI). All experiments were performed in triplicate

Fig 5: High plasma levels of Hcy are closely related to the occurrence of cardiac fibrosis. A‐B, The expression levels of TRPC3 and the relevant proteins, such as TGF‐β and Collagen‐I in AF patients. C, The relative mRNA level. D1‐D2, The atrial nucleus in patients with AF is shown with haematoxylin and eosin (HE) staining, Masson's trichrome ‐stain and immunohistochemistry, as well as statistical results of positive areas and ratio to total area of the fibre (Magnification × 100, Scale bar = 200 μm; n = 4 per group). E, The ultra‐structure of atrial tissues was observed by electron microscopy: E1‐E3: SR patients, E4‐E6: AF patients. Error line indicates mean and standard deviation. *P < .01, **P < .001. Multiple t‐tests were performed using step‐down bootstrap sampling to control the familywise error rate at 0.05 for 1B, C, and unpaired t‐test was used for 1D. Fisher's exact test was used for analysing Table 1