Fig. 2.

Decreased NFκB transcription is associated with reduced MGMT protein levels. MGMT protein levels after combination BTZ+TMZ treatment in (a) P3 and (b) T98G cell lines. GAPDH was used as a loading control. Band intensity expressed as a ratio after normalisation to GAPDH. (c) Chymotryptic activity of the β1 and β5 subunits measured as relative fluorescence units (RFU/380/460 nm) of fluorophore 7-amino-4-methylcoumarin in cells treated as indicated. (d) Fold change in transcription factor activity in nuclear extracts from P3 cells. NFκB/p65 subunit levels in P3 cell extracts from (e) total protein, and (f) nuclear fraction after treatment, as indicated. (g) Representative peak heights and nucleotide disposition order (NDO) within MGMT promoter CpG region (h) % MGMT promoter methylation in P3 cells after treatment. Each experiment was performed in triplicate, and the data represent the mean ± S.E.M. of at least three independent experiments, *P < 0.05 and ****P < 0.0001