NFκB expression in medulloblastoma in vitro and in vivo. A: Activation of NFκB: Timecourse of NFκB activation by 15 ng/mL TNFα in Daoy cells by probing for nuclear p65, nuclear phos(S276) p65, & cytoplasmic IκB. nuc = nuclear fraction, cyto = cytoplasmic fraction. p65 band ~70kD; phos p65 band ~97kD; IκB band ~37kD B: Tissue Expression: expression of NFκB p65 subunit in D425 cells grown in culture (CC) compared to D425 cells grown in a flank tumor model (FT). Nuclear (nuc) and cytoplasmic (cyto) fractions were separated by differential centrifugation following lysis by nitrogen cavitation as described. C: p65 expression by Western in cultured cells vs flank tumors for 2 additional cell lines. CC = cells in culture, FT = flank tumor. p65 band~60kD. Extracts were obtained using the NE-PER kit as described.