Fig. 2: CB₁ hotspots are connected with components of the MPS.

a Two-color STED images of CB₁ (magenta) and βII-spectrin (green) in the axons of cultured neurons. N = 3 biological replicates. b, c Top, enlarged images taken from white boxes from (a). 1D projection traces of βII-spectrin (green) and CB₁ (magenta) signals along the axon are shown in the middle. 1D cross-correlation functions between the distributions of CB₁ and βII-spectrin from CB₁-positive axon segments are shown in the bottom. d–f PLA was performed in cultured neurons of WT mouse (d), cnr1^−/− mouse (e), and tetracycline-induced CB₁ transfected CHO cells (f) with antibody of CB₁ and ankB. Cell nuclei were stained with DAPI (blue). N = 3 biological replicates. g Immunoprecipitation of CB₁ and ankB in CB₁-CHO cells. Samples were processed for immunoprecipitation with either anti-CB₁ or IgG control antibodies. Immunoprecipitates were immunoblotted with the anti-CB₁ antibody (55kD) and anti-ankB antibody (220kD). N = 2 biological replicates. h PLA was performed in HEK-293T cells coexpressing ankB and different fragments of CB₁, including wild type (CB1), CB₁ with truncated ICL3 loop (CB₁ΔICL3), or CB₁ with truncated C-terminal (CB₁ΔC-term). ratio r from left to right: 14.1 ± 0.5, 6.3 ± 0.4, 5.5 ± 0.4. Data are mean ± s.e.m. (N = 3 biological replicates; 25–50 imaging regions were examined per condition). ***p < 0.0001, statistical analysis was performed by unpaired two-tailed Student’s t test. Source data are provided as a Source Data file.