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. 2017 Aug 24;8(56):95256–95269. doi: 10.18632/oncotarget.20438

Figure 2. TCTN2 is expressed in cancer cell lines and localizes in the inner side of plasma membrane.

Figure 2

(A) Total proteins were extracted from breast (SKBR3 and MCF7), lung (H226 and HOP92), colon (HCT15, HT29 and Colo205) and ovarian (SKOV3 and OVCAR8) cancer cell lines. Proteins were resolved by SDS-polyacrylamide gel electrophoresis and analyzed by Western blot with anti-TCTN2 monoclonal antibody. Anti-β-Actin monoclonal antibody was used as a loading control. (B) TCTN2 was silenced in HOP92, OVCAR8 and HCT15 cancer cell lines with four commercially available (QIAGEN) TCTN2-specific siRNAs at 10 nM concentration or irrelevant siRNA (AllStars Negative Control siRNA, QIAGEN) using the Hi-Perfect transfection reagent (QIAGEN). TCTN2 expression was assayed 48 hours later by Western blot. Anti-β-Actin monoclonal antibody was used as a loading control. (C) TCTN2 localizes in the inner side of plasma membrane in cancer cells lines. HCT15, HOP92 and HT29 cells were analyzed by confocal microscopy. Cells were incubated with the anti-TCTN2 monoclonal antibody, with (lower panels) or without (upper panels) permeabilization pre-treatment with 0.01% BriJ96®. Cells were subsequently stained with Alexafluor 488-labeled goat anti-mouse antibodies to detect TCTN2 (green) and DAPI to visualize nuclei (blue).