Figure 6. Rab2a recruits HOPS to LEs.
(A–D) PNSs and FF endosomes (20/120 min [pulse/chase]) from mock-treated or Rab2a KD cells were analyzed by immunoblotting. Representative immunoblots quantified in (B)–(D).
(E) PNSs, membrane (mem), and soluble protein (cyt) fractions from mock-treated or Rab2a KD cells were analyzed by immunoblotting.
(F–I) Immunoblot analysis of different-aged FF endosomes from HeLa WT cells; quantification in (G)–(I). Protein levels in 120 min samples were set as 100%.
(J) PNSs from HeLa WT or Arl8a/b KO transfected with non- or Rab2a-targeting siRNAs were fractionated into membrane-bound and cytosolic proteins as in (E).
(K–N) HeLa cells were treated with 1 μg/mL BFA for 30 min at 37°C. Purified FF endosomes (30/120 min [pulse/chase]) were analyzed by immunoblotting. BFA was present throughout the incubations.
(K) Representative immunoblots quantified in (L)–(N).
Data are means and SDs from 3 independent experiments (B–D, G–I, and L–N).