Primary antibodies were: mouse anti-human HSP70, specific for stress-inducible HSPA1A (StressMarq, Canada, SMC-100B; 1:100 ICC; 1:1000 WB),
mouse anti-Flag M2 (Sigma-Aldrich, #F1804; 1:400 ICC); rabbit anti-FUS (Proteintech, USA, 11570-1-AP; 1:400 ICC), mouse anti-human SOD1 (Sigma-Aldrich Canada, SD-G6; 1:100 ICC), rabbit anti-acetyl-histone H3K9/K14 (Cell Signaling, #9677; 1:400 ICC; 1:1000 WB), mouse anti-GAPDH (MediMabs, Canada, #MM-0163; 1:1000 WB), moue monoclonal anti-acetylated tubulin (Sigma-Aldrich #T6793; 1:1000 WB) and
rabbit anti-α-tubulin (Abcam #ab15246; 1:1000 WB).
Secondary antibodies (Jackson Immunoresearch: Cedarlane, Canada):
Alexa Fluor 488-conjugated Affinipure donkey anti-mouse IgG (1:300);
Cy3-conjugated donkey anti-mouse IgG 1:300);
Cy5-conjugated donkey anti-rabbit IgG (1:300), and
HRP-conjugated goat anti-mouse (1/5000 for WB, 1/500 for IC) or
donkey anti-rabbit IgG (Jackson Immunoresearch (1:2500).
HDAC inhibitors and HSP-inducing drugs: SAHA (suberoylanilide hydroxamic acid) and
tubastatin A (Cayman Chemical: Cedarlane, Canada);
tacedinaline,
RGFP109 and
RGFP966 (Selleckchem: Cedarlane, Canada); sodium phenylbutyrate (Selleckchem); arimoclomol (Toronto Research Chemicals, Canada); NXD30001 was previously supplied by NexGenix Pharmaceuticals (Cha et al. 2014 (
link)).
Kuta R., Larochelle N., Fernandez M., Pal A., Minotti S., Tibshirani M., St. Louis K., Gentil B.J., Nalbantoglu J.N., Hermann A, & Durham H.D. (2020). Depending on the stress, histone deacetylase inhibitors act as heat shock protein co-inducers in motor neurons and potentiate arimoclomol, exerting neuroprotection through multiple mechanisms in ALS models. Cell Stress & Chaperones, 25(1), 173-191.