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| Figure 7: shRNA mediated knockdown of LYPD6B. Four different lentiviral shRNA particles of LYPD6B were transduced to OVCAR 3 cell. (A) qPCR was utilized to quantitate levels of LYPD6B in knockdown cells using assay Hs00962009_m1 that detects all variants. Knockdown efficiency of shLYPD6B #1, #3, #4 and #5 was 43, 90, 78 and 52 % compared to the parental cell LYPD6B expression levels, respectively. (B) LYPD6B immunoblot result of knockdown cell. Four different LYPD6B knockdown cells were lysed by RIPA buffer and equal amount of whole cell proteins were analyzed by immunoblot using anti-LYPD6B antibody. LYPD6B protein levels were significantly decreased in knockdown cells compared to the parental (WT) and nontarget shRNA transducted (shNT) cell. (C) Proliferation assay of LYPD6B knockdown cell. 2000 cell of WT, shNT and four shLYPD6B were seeded to 96 well p late and cultured for 5 days and viable cells were measured by MTS assay. There were no differences of cell proliferation in these LYPD6B knockdown cells. |
