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. 2001 Aug 1;108(3):425–435. doi: 10.1172/JCI12629

Figure 2.

Figure 2

SDF-1α induction of NF-κB activity is indirect but correlates with degradation of IκB-α and IκB-β. (a) SDF-1α induction of NF-κB activity is slow but correlates with degradation of IκB-α and IκB-β. Murine primary astrocytes were incubated with medium alone or with SDF-1α or TNF-α for the indicated periods of time. Whole-cell extracts were then prepared and subjected to NF-κB DNA binding activity assays by EMSA with IP-10 κB2 probe (top panel: SDF-1α treatment; bottom panel: TNF-α treatment). Additional aliquots of the same whole-cell extracts were subjected to anti–IκB-α and anti–IκB-β Western blot (middle panel). After stripping, the same membrane was blotted with anti-STAT3 (middle panel) to monitor protein loading for each sample. Data represent three experiments. (b) CHX suppresses SDF-1α–induced but not TNF-α–induced NF-κB activation. Murine primary astrocytes were treated with SDF-1α (lane 2) for 2 hours, TNF-α (lane 5) for 30 minutes, CHX (50 μg/ml, lane 3) for 1 hour, or CHX for 1 hour followed by either SDF-1α (lane 4) for 2 hours or TNF-α (lane 6) for 30 minutes. Whole-cell extracts were then prepared and analyzed by EMSA with the IP-10 κB2 probe. Data represent three experiments.