Plan apochromat x 63 na 1.4 oil immersion objective

Confocal microscope analysis was performed as previously described [38 (link)]. Briefly, cells were fixed with 3% paraformaldehyde and permeabilized with 0.1% Triton X-100 before incubation with specific antibodies against NF-kB (1:100, rabbit) and Vimentin (1:1.000, mouse). Secondary antibodies were Alexa Fluor 488 (1:1000) or Alexa Fluor 633 (1:1000). Microscopy analyses were performed using a Zeiss LSM 700 confocal microscope (Carl Zeiss MicroImaging, Jena, Germany) equipped with a plan-apochromat X 63 (NA 1.4) oil immersion objective (Zeiss, Thornwood, NY, USA).

Confocal laser scanning microscopy analysis were performed as previously described^{13 (link)}. LoVo cells (8 × 10³/well) were seeded in 24-well plate containing microscope glass. After treatment, cells were fixed using 4% (v/v) paraformaldehyde solution for 20 min and then permeabilized with 0.1% (v/v) Triton X-100 in PBS for 10 min at RT. SIRT6 immunofluorescence detection was performed by using specific antibodies against Actin (1:1000) and SIRT6 (1:500). Alexa Fluor 488 (1:1000) or Alexa Fluor 633 (1:1000) were used as secondary antibodies. Zeiss LSM 700 confocal microscope with a plan apochromat X63 (NA1.4) oil immersion objective was utilized to perform microscopy analyses, as previously described^{13 (link)}. The fluorescence intensity was estimated with ImageJ software 1.52n version and expressed as arbitrary fluorescence units (AFU).