Anti rap1

Western blot was done with a modified version of a previous method [24 (link)]. The protein expression levels were calculated as folds of normal liver or untreated cells. β-Actin, anti-NF-κB, anti-Rap1, anti-JNK, anti-ERK and anti-p38 antibodies were purchased from Cell Signaling Technology or ABCAM.

Proteins were separated by SDS-PAGE (10–15%) and transferred to a PVDF membrane, which were incubated (overnight, 4°C) with antibodies: anti-EPAC1 (1/500), anti-caspase 3 (1/500), anti-caspase 9 (1/1000), anti-RAP1 (1/1000), and anti-H₂AX-pS₁₃₉ (1/1000) from Cell Signaling Technology (Saint-Cyr-L’Ecole, France), anti-TopIIβ (1/1000) from Abcam (Paris, France), anti-SERCA2A (1/1000), and anti-actin (1/50,000) from Santa Cruz Biotechnology (Heidelberg, Germany), anti-calsequestrin (1/2500) from Thermo Fisher Scientific (Les Ulis, France), and anti-tubulin (1/1000) from Sigma (St Quentin Fallavier, France). Proteins were detected on the iBright FL1000 Imager (Thermo Fisher Scientific, Les Ulis, France) with ECL and protein band intensity was calculated using ImageJ software and normalized by actin.

Proteins were separated by SDS-PAGE (10–15%) and transferred to a PVDF membrane, which were incubated (overnight, 4°C) with antibodies: anti-EPAC1 (1/500), anti-caspase 3 (1/500), anti-caspase 9 (1/1000), anti-RAP1 (1/1000), and anti-H₂AX-pS₁₃₉ (1/1000) from Cell Signaling Technology (Saint-Cyr-L’Ecole, France), anti-TopIIβ (1/1000) from Abcam (Paris, France), anti-SERCA2A (1/1000), and anti-actin (1/50,000) from Santa Cruz Biotechnology (Heidelberg, Germany), anti-calsequestrin (1/2500) from Thermo Fisher Scientific (Les Ulis, France), and anti-tubulin (1/1000) from Sigma (St Quentin Fallavier, France). Proteins were detected on the iBright FL1000 Imager (Thermo Fisher Scientific, Les Ulis, France) with ECL and protein band intensity was calculated using ImageJ software and normalized by actin.