Anti human caspase 1

The hPDLSCs-CTR and hPDLSCs-MOR were processed as previously reported by Trubiani et al. (2012) [88 (link)]. Primary monoclonal antibodies anti-human Caspase 1 (1:200, rabbit) (Abcam, Milan, Italy), anti-human SOD1 (1:200, rabbit) (Abcam), anti-human Bax (1:100, rabbit) (Cell Signaling Technology, Milan, Italy), anti-human Bcl2 (1:200, rabbit) (Cell Signaling Technollogy, Milan, Italy), and anti-human LC3A/B (1:100, rabbit) (Cell Signaling Technology) were used, followed by Alexa Fluor 568 conjugated goat anti rabbit as secondary antibodies (1:200; ThermoFisher, Life Tech., Monza, MB, Italy) for 1 hr at 37 °C. Subsequently cells were incubated with AlexaFluor 488 phalloidin green fluorescence conjugate (1:200; ThermoFisher, Life Tech.), to evidence cytoskeleton actin. Cell nuclei were stained with TOPRO (1:200; ThermoFisher) for 1 hr at 37 °C. Glass coverslips were placed face down on glass slides and mounted with Prolong antifade (ThermoFisher, Life Tech.) [91 (link)]. Samples were observed by means of a Zeiss LSM800 confocal system, connected to an inverted Zeiss Axiovert 200 microscope equipped with a Plan Neofluar oil-immersion objective. Images were collected using an argon laser beam with excitation lines at 488 nm and a helium-neon source at 543 and 633 nm. Post-acquisition image analyses were carried out with a Zeiss ZEN software.