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. 2021 Feb 8;118(7):e2017937118. doi: 10.1073/pnas.2017937118

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Copyright © 2021 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 2. — Rab4a and Rab11a regulate an ISO-stimulated boost in Ca_V1.2 recycling. (A) TIRF images of Ca_v1.2-tRFP distribution in the PM of tsA-201 cells before (Left) and after 100 nM ISO (Right; n = 18). (B) Time course and kinetics (solid red line) of the fold-change in Ca_V1.2-tRFP intensity in the TIRF footprint before, during, and after ISO stimulation. (C) Histogram summarizing Ca_V1.2 channel cluster density (number/μm²) in the TIRF footprint in control and ISO-stimulated conditions. (D–F) Same layout format in tsA-201 cells coexpressing constitutively active (GTP-locked) Rab4a (n = 10). (G–I) Same layout format in tsA-201 cells coexpressing dominant-negative (GDP-locked) Rab4a (n = 12). (J–L) Same layout format in tsA-201 cells coexpressing dominant-negative (GDP-locked) Rab11a (n = 8). (M) Calculation and resultant theoretical time-course and kinetics of Rab11a-dependent ISO-stimulated recycling. Data that passed a normality test (C and L) were analyzed with a paired t test, others (F and I) were analyzed with a nonparametric Wilcoxon test. ***P < 0.001; **P < 0.01; *P < 0.05.