An optimized Bcl-3 inhibitor for melanoma treatment

Fig. 1F
Left panel: Lysates prepared from Mel Ho and B16F10 cells treated with different concentrations A27 or DMSO for 24 hours followed by Western Blot analysis toward Bcl-3 cyclin D1 (CD1), and α-Tubulin.
Right panel: Lysates prepared from HT144 cells treated with different concentrations A27 or DMSO for 24 hours followed by Western Blot analysis toward Bcl-3, cyclin D1, and α-Tubulin.

Fig. 2F
Overexpression of FLAG-Bcl-3 in Mel Juso cells that lack Bcl-3 expression followed by Flag-immunoprecipitation in cells treated with or without A27 for 24 hours, analyzed with an anti-Flag-, anti-p50, and anti-actin antibody.

Suppl. Fig. 2B
In vitro pull-down assay using empty-GST, GST-Bcl-3, and His-p50 antibodies incubated with or without 50 μM or 200 μM of A27 for 1h, followed by separation and elution by μMACS columns. Eluates were subjected to Western Blot analysis toward p50 and GST.

Suppl. Fig. 2C
Mel ju cells transiently transfected with ON-TARGETplus SMARTpool of siRNA targeting BCL-3 (25nM, Dharmacon), or ON-TARGETplus Non-targeting Pool (siControl) (25nM, Dharmacon) using DharmaFECT Transfection reagent (Dharmacon) for 48 hours before cell lysate and western blotting against Bcl-3 or actin.

Suppl. Fig. 4C
Western blot using Cyclin D1 or actin in tumor lysate isolated from vehicle control or A27-treated mice.