Figure 2.

Surface oligomerization is preserved with deletion of the distal Panx1CT. (a) Crosslinking assays reveal oligomerization profiles of C-terminal deletion mutants. Representative Western blots observing HEK293T cell lysates post-transfection with (i) Panx1-EGFP, (ii) Panx1∆379-EGFP, or (iii) Panx1∆299-EGFP, and after crosslinking with the cell-impermeable crosslinker, BS³, to observe surface-localized oligomers (6× or ~2–3×). The plot of each quantification is included to shed light on the analytical process. An antibody for Crmp2, an intracellular protein known to form tetramers, was used as a negative control to ensure BS³ had not entered the cell. Crmp2 oligomerization did not increase in the presence of BS³, as expected. (b) Each oligomeric band was quantified and expressed as a percentage of the entire GFP signal in each lane. Non-specific bands were excluded from the analysis. Mutants lacking the distal Panx1CT (Panx1∆379-EGFP) had the same oligomerization profiles as full length Panx1-EGFP. Data are presented as mean ± SEM. One-way ANOVA with Dunnett’s multiple comparisons test, N = 3, α = 0.05; F(2,6) = 17.59, **P = 0.0023 (6×); F(2,6) = 15.73, *P = 0.0127 (3×); F(2,6) = 13.84, *P = 0.0169 (1×); ns, non-significant. All samples were derived from the same experiment and processed in parallel. This figure was modified from Epp 2019⁴⁶. For uncropped images of all Western blots in this figure, please see Supplementary Fig. S4