Supplementary materials for the study "TRPV3 activation by different agonists accompanied by lipid dissociation from the vanilloid site" by Kirill D. Nadezhdin et al. include figures and tables. The figures present cryo-EM data processing workflows for TRPV3 in different states (APO, OPEN, INACT) with various agonists (THCV, 2-APB). Figure S1 shows the workflow for the APO structure, including micrographs, 2D class averages, and 3D classes. Figure S2 provides characteristics of cryo-EM reconstructions for different TRPV3 structures. Figure S3 shows the workflow for TRPV3 in the presence of THCV. Figure S4 compares pore structures in open-state TRPV3 structures and shows pore radii calculated using HOLE. Figure S5 shows the workflow for TRPV3 in the presence of 2-APB. Figure S6 presents cryo-EM density of TRPV3, showing fragments of the TMD and close-up views of the S1-S4 base site. Figure S7 shows 2-APB-bound open and inactivated states, including pore-forming domains, S6 bundle crossings, and interfaces between neighboring subunits. Tables S1 and S2 provide cryo-EM data collection, refinement, and validation statistics, as well as EC50 values for TRPV3 activation by THCV and 2-APB. The data highlights structural changes in TRPV3 upon agonist binding and the role of lipid dissociation in vanilloid site activation.Supplementary materials for the study "TRPV3 activation by different agonists accompanied by lipid dissociation from the vanilloid site" by Kirill D. Nadezhdin et al. include figures and tables. The figures present cryo-EM data processing workflows for TRPV3 in different states (APO, OPEN, INACT) with various agonists (THCV, 2-APB). Figure S1 shows the workflow for the APO structure, including micrographs, 2D class averages, and 3D classes. Figure S2 provides characteristics of cryo-EM reconstructions for different TRPV3 structures. Figure S3 shows the workflow for TRPV3 in the presence of THCV. Figure S4 compares pore structures in open-state TRPV3 structures and shows pore radii calculated using HOLE. Figure S5 shows the workflow for TRPV3 in the presence of 2-APB. Figure S6 presents cryo-EM density of TRPV3, showing fragments of the TMD and close-up views of the S1-S4 base site. Figure S7 shows 2-APB-bound open and inactivated states, including pore-forming domains, S6 bundle crossings, and interfaces between neighboring subunits. Tables S1 and S2 provide cryo-EM data collection, refinement, and validation statistics, as well as EC50 values for TRPV3 activation by THCV and 2-APB. The data highlights structural changes in TRPV3 upon agonist binding and the role of lipid dissociation in vanilloid site activation.