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. 2011 Apr 14;6(4):e18762. doi: 10.1371/journal.pone.0018762

Figure 6. Knockdown of NET4/TMEM53 in U2OS and MRC5 cells.

Figure 6

(A) Splice variants and position of knockdown oligos. (B) NET4/TMEM53 transcript levels were effectively knocked down in MRC5 cells using both siRNA oligos and an esiRNA. Peptidylprolyl isomerase A (PPIA) was used as a control for normalization. (C) NET4/TMEM53 transcript levels were also knocked down in the U2OS cell line using the siRNA oligo TMEM53 si2. The scramble siRNA oligo had no effect. PPIA was used as a control for normalization. (D) Western blot demonstrating the knockdown of NET4/Tmem53. Because antibodies to NET4/Tmem53 recognized multiple bands on Western blot at the expected molecular weight, knockdown of the protein was tested using a NET4/Tmem53-GFP fusion and GFP antibodies. The siRNAs were transfected 24 h after transfection of NET4/Tmem53-GFP in order to enable protein to be generated from the plasmid prior to the beginning of knockdown.