A, ALS marker proteins HSP27, 20S proteasome, and ubiquitinated proteins are co-localized with tubulin aggregates in BITC-treated cells.HeLa cells were treated with 5 μM BITC for 1 h before being fixed and immunofluorescent-stained for ALS markers (in red) and α-tubulin (in green). B, ITC treatment induces rapid binding between tubulin and ALS marker proteins. HeLa cells were treated with 10 μM BITC for up to 1 h. The insoluble fraction was extracted and immunoprecipitated for α-tubulin. The IP products were immunoblotted for ALS markers. C, the potency order of inducing ALS formation is: BITC>PEITC>SFN. HeLa cells were treated with 10 μM BITC for 1 h. The insoluble fraction was extracted and immunoblotted for α-tubulin and ALS markers.