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Fig. 8. Effects of kinectin RNAi on anterograde distribution of ER and mitochondria in HeLa cells. (A) mRNA levels in untransfected cells (lane 1), vector alone stable cells (lane 2) and kinectin knockdown stable cells (lane 3) were determined by semi-quantitative RT-PCR. (B) 10 µg of whole cell lysates as in (A) subjected to western blotting for kinectin and actin proteins. (C) Silencing of kinectin protein. Untransfected HeLa cells (a1-4) and HeLa cells transfected with pSilencer vector control siRNA (b1-4), or with kinectin siRNA (c1-4). Kinectin knockdown stable cells showed distinct disruption in kinectin (c1), ER (c2) and mitochondria (c3) distribution. By contrast, vector control cells (b1-3) and untransfected cells (a1-3) show normal staining patterns. Each cell type was stained for kinectin (anti-CT-1) (red); ER (anti-calreticulin) (red); mitochondria (anti-Mn SOD) (red) and DNA (DAPI) (blue). Microtubule (anti-ß-tubulin) (red) distribution appeared normal for all cells type (a4-c4). Bars, 20 µm.





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