Figure 3.
Figure 3. Absence of ARH leads to apical missorting of LDLR-CT27. (A) Lysates of HBE cells stably depleted of ARH or GAPDH were analyzed by SDS-PAGE and quantitative Western blotting. Data represent mean values of three independent experiments, each with three different data points representing 2.5, 5, and 7.5 µg total protein. Errors are SD. (B–E) Knockdown cells were grown on filter supports and infected with defective adenoviruses expressing LDLR-CT27 (B), LDLR(Y18A) (C), FcR-RFP (D), or NgCAM-CT43 (E). Receptors at the surface were stained using cargo-specific antibodies. GFP expression indicates shRNA presence. Specimens were analyzed by confocal microscopy and representative images are shown. Data mentioned in the text were from three independent experiments scoring 30 Arf6 and 22 GAPDH knockdown cells for randomized apical and basolateral sorting of LDLR-CT27. Bars, 5 µm.

Absence of ARH leads to apical missorting of LDLR-CT27. (A) Lysates of HBE cells stably depleted of ARH or GAPDH were analyzed by SDS-PAGE and quantitative Western blotting. Data represent mean values of three independent experiments, each with three different data points representing 2.5, 5, and 7.5 µg total protein. Errors are SD. (B–E) Knockdown cells were grown on filter supports and infected with defective adenoviruses expressing LDLR-CT27 (B), LDLR(Y18A) (C), FcR-RFP (D), or NgCAM-CT43 (E). Receptors at the surface were stained using cargo-specific antibodies. GFP expression indicates shRNA presence. Specimens were analyzed by confocal microscopy and representative images are shown. Data mentioned in the text were from three independent experiments scoring 30 Arf6 and 22 GAPDH knockdown cells for randomized apical and basolateral sorting of LDLR-CT27. Bars, 5 µm.

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