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. 2014 Nov;34(21):3981–3992. doi: 10.1128/MCB.00113-14

FIG 6.

FIG 6

Impact of knockout expression of Tpcn1A/B or Tpcn2 in cholera toxin trafficking. (A) Schematic representation of CTxB trafficking from the plasma membrane to the Golgi apparatus. (B) Representative images for 0-min (no chase) and 120-min chase periods at 37°C, after 30 min incubation on ice with Alexa Fluor 555-labeled CTxB. Cells were immunolabeled with an antibody against the Golgi apparatus marker protein GM130. Cell boundaries are identified with a white broken line. (C) Quantification of CTxB levels in the Golgi apparatus at the indicated chase time after the CTxB binding period (n = 56 to 93). (D, E) Effects of treatment of WT MEFs with shRNA (shRNA targeting Rab7b or a scrambled sequence control) on relative Rab7b mRNA levels determined by RT-qPCR (D) and on CTxB levels in the Golgi apparatus after a chase period of 120 min (E) (n = 32 to 36). Data points correspond to the mean ± SEM. ns, no significant difference (P > 0.05); **, P < 0.01; ***, P < 0.001. Primary MEFs (passage numbers, <5) were used in all experiments.