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Fig. 2 | Cell & Bioscience

Fig. 2

From: Subcellular fractionation method to study endosomal trafficking of Kaposi’s sarcoma-associated herpesvirus

Fig. 2

Detection of KSHV in endosomal fractions. A Distribution of endosomal membranes was determined from uninfected and KSHV infected HFF cells using sucrose gradient fractions. Equal-volume aliquots from each gradient fraction were separated by SDS-PAGE and probed by Western blotting against organelle markers: Rab5 and EEA1 (EE markers), Rab7 (LE marker), LAMP1 (LE and lysosome marker), and histone H3 (nuclear marker). B Lysosome detection in sucrose gradient fractions of uninfected and KSHV infected HFF cells. Acid phosphatase activity was analyzed by using Acid Phosphatase Assay Kit (Sigma). C KSHV escapes from the LE. Following gradient centrifugation, viral genomic DNA was extracted from each recovered fraction. Fractions were then quantified for the content of internalized viral DNA by performing qPCR using specific primers. Data (panels B, C) represent the average ±SD (error bars) of three experiments. Columns with different alphabets indicate statistical significance (p < 0.05) by least significant difference (LSD)

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