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. 2021 Apr 30;7(18):eabf3873. doi: 10.1126/sciadv.abf3873

Fig. 4. Endolysosomal sorting of VAMP4 regulates its abundance in the SV pool.

Fig. 4

(A and B) Hippocampal neurons cotransfected with VAMP4-pHluorin and either wild-type (WT), constitutively active (Q67L), or dominant-negative (T22N) mCherry-rab7. Scale bar, 5 μm. Representative images (A) and bar graph (B) display synaptic VAMP4-pHluorin expression. n = 23 (mCherry), n = 12 (mCherry-rab7), n = 13 (Q67L), and n = 18 (T22N) coverslips from four independent preparations, ****P < 0.001, Kruskal-Wallis test with Dunn’s multiple comparisons test. (C) Effect of T22N mCherry-rab7 overexpression on sypHy synaptic expression. n = 21 (mCherry) or n = 16 (T22N mCherry-rab7) coverslips from four independent preparations, ***P < 0.0002, Mann-Whitney test. (D) Fusion events reported by VAMP4-pHluorin upon stimulation with 400 APs at 40 Hz under the conditions shown in (A) and (B), n = 23 (mCherry), n = 12 (mCherry-rab7), n = 13 (Q67L), and n = 18 (T22N) coverslips from four independent preparations, *P = 0.0129, Kruskal-Wallis test with Dunn’s multiple comparisons test.