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. 2012 Dec 14;7(12):e52060. doi: 10.1371/journal.pone.0052060

Figure 6. gp120 and/or MA activates IκBα and increases the translocation of p50 in SVGA astrocytes.

Figure 6

(A) SVGA astrocytes were treated with 500 μM MA and/or 50 ng/ml recombinant gp120 protein for 0, 5, 10, 20, 40 and 60 minutes and the whole cell lysate was prepared by lysing the cells with RIPA buffer. The proteins were separated using SDS-PAGE electrophoresis as mentioned in the Materials and Methods section. The p-IκBα was determined in western blotting. GAPDH was used as housekeeping control. (B) SVGA astrocytes were treated with MA and/or transfected with gp120 for 6 hours. The translocation of p50 was measured from cytoplasm to nucleus as described in the Materials and methods. GAPDH was again used as loading control for cytoplasmic extracts and LaminB was used as loading control for nuclear extracts. The bar charts below the representative images show the densitometry values corresponding to the respective treatments. The housekeeping proteins were used to normalize the protein expressions of various genes.