Fig. 5

NFκB p65 activator compounds cause p65 nuclear translocation in NPCs and reverse the proliferation deficit in case-derived NPCs. a Structures of NFκB p65 activators compound 1 (SRI-22772) and compound 2 (SRI-22782). b NPCs (from controls and cases) were plated down in default media (without mitogens) and 7–10 days later TNF-α or compounds 1 or 2 were added to the media for 24 h. c NPCs treated with either TNF-α (positive control) or compounds 1 or 2 showed significantly increased anti-phospho-NFκB p65 (Ser536) localising to the nucleus compared to vehicle-treated. d Nuclear extracts were prepared from NPCs treated with vehicle (DMSO control), TNF-α and compounds 1 and 2 and immunoblotted with anti-phospho-NFκB p65 (Ser536). e Bands were quantified and normalised to the levels of vinculin (loading control). Compound 2 treatment resulted in significant increase in nuclear NFκB p65 Ser536 expression (ordinary one-way ANOVA. Values plotted are the mean  ± sem; *p < 0.05; ns non-significant; ****p < 0.0001). f Representative images of proliferation assays of case NPCs (case 1, clone 1) following treatment with vehicle (DMSO), TNF-α, compounds 1 and 2. g Graph showing proliferation of all case NPCs (pooled) showing significant rescue of proliferation deficits following treatment with TNF-α and compounds 1 and 2 (ordinary one-way ANOVA with Dunnett’s multiple comparison test. Values plotted are the mean  ± sem; ****p < 0.0001). Representative images of case NPCs infected with NFκB p65 (RELA) Lentivirus or vector alone (control) demonstrate that cells infected with RELA express increased NFκB p65 and exhibit increased proliferation i compared to NPCs from the same patient infected with the vector alone