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. Author manuscript; available in PMC: 2016 Apr 12.
Published in final edited form as: Cell Rep. 2016 Mar 22;14(12):2962–2974. doi: 10.1016/j.celrep.2016.02.068

Figure 2. α3 regulates α4-α4 proteasome assembly in human cells.

Figure 2

(A) Cartoon illustrating regions of the α3 gene targeted by different shRNAs.

(B) shRNA-mediated knockdown of α3 in stable α4-CC-Flag (HeLa) cells. The α3-1 shRNA cause a ∼50% decrease in α3 levels. α4-CC-Flag expression was induced by the addition of doxycycline (Dox).

(C) α3 knockdown in α4-CC-Flag stable HeLa cells results in increased assembly of α4-α4 proteasomes. Also see Fig. S2.

(D) Confirmation of the increase in α4-α4 proteasomes upon α3 knockdown using gel-isolated HeLa cell 26S proteasomes.

(E) Ectopic overexpression of α3 in α4-overexpressing HeLa cells causes a decrease in α4-α4 proteasome formation based on α4- α4 crosslinking.

(F) α6-Flag expression was induced in stable α6-Flag HeLa cells by addition of Dox; α4-WT or α4-CC was transiently expressed in these cells.

(G) α6 expression level does not affect α4-α4 proteasome assembly. α6-Flag expression was induced in α6-Flag stable HeLa cells by addition of Dox (see Fig. 2F).