Bs 7651r

Separate staining for DNase1 and DNase1L3 was performed using the protocol described previously [73 (link)]. Sections were incubated overnight at 4 °C with DNase1-polyclonal antibody (BS-7651R, Bioss, Woburn, MA, USA, 0.01 µg/mL) or DNase1L3-polyclonal antibody (BS-7653R, Bioss, Woburn, MA, USA, 0.01 µg/mL). The next day, sections were incubated for 30 min at room temperature with a donkey anti-rabbit-IgG Alexa Fluor 647 (AB150075, Abcam, Cambridge, UK) and DAPI (Invitrogen, Grand Island, NY, USA). Images were captured using a fluorescence microscope (KEYENCE BZ-9000, Keyence Corporation, Osaka, Japan). Exposure time was calibrated using the isotype control antibody (AB37415, Abcam, Cambridge, UK), with adjustments made to achieve a negative fluorescence signal in the control. Three to five samples were inspected microscopically, and one representative image was taken per condition.

A separate staining for DNase1 and DNase1L3 was performed using the protocol described above. As primary antibodies a DNase-1-polyclonal-antibody (BS-7651R, Bioss, Woburn, MA, USA) and a DNase-1L3-polyclonal-antibody (BS-7653R, Bioss, Woburn, MA, USA) were used in a concentration of 0.01 µg/ml. Rabbit-IgG (AB37415, Abcam, Cambridge, UK) served as isotype control. As secondary antibody, a donkey-anti-rabbit-IgG Alexa Fluor 647 (AB150075, Abcam, Cambridge, UK) was used.