Mouse anti cdh1

Immunoblot analysis was performed as previously described [23 (link)], using rabbit anti-TFF3 antibody [14 (link)]. Mouse anti-β-ACTIN, rabbit anti-p-c-SRC, mouse anti-c-SRC, and mouse anti-γ-CTNNG antibody was obtained from Santa Cruz Biotechnology, Santa Cruz, CA, USA. Mouse anti-CDH1, mouse anti-CDH2, rabbit anti-OCLN, mouse anti-VIM, mouse anti- ITGA6, rabbit anti-pSTAT3, and mouse anti-STAT3 antibody was obtained from Abcam, Cambridge, MA, USA. Cell extracts were resolved by SDS-PAGE and immunoblotted, with the respective antibodies, as previously described [23 (link)]. β-ACTIN was used as input control for cell lysate. The sizes of detected protein bands in kDa are shown on the left side.
Confocal laser scanning microscopy was performed as previously described [22 (link)]. Nuclei were visualised with VECTASHIELD mounting medium with DAPI (Vector Laboratories, Burlingame, CA, USA). Images were captured using a confocal microscope (Eclipse C1 Plus Confocal; Nikon Instrument Inc., Melville, NY, USA). Primary antibody, Mouse anti-CDH1, and mouse anti-VIM was obtained from Abcam. Secondary antibody, Alexa Fluor 488 rabbit anti-mouse immunoglobulin (Ig)G and Alexa Fluor 568 rabbit anti-mouse IgG was obtained from Invitrogen, Singapore. Rhodamine-conjugated phalloidin (Sigma-Aldrich, St Louis, MO, USA) was used to visualize f-actin filaments.

Free floating sections were washed in Tris buffered saline with Triton solution (0.1%) (TBS-Tx), 3 times at room temperature (RT). Sections were incubated for 3 h at RT in a pre-incubation solution of 2% bovine serum (BSA; A7906, Sigma) in TBS-Tx. After the pre-incubation, the tissue was incubated, 2 days at 4 °C, in a primary solution containing BSA (2%) in TBS-Tx with the primary antibodies: rabbit anti-Glutaminase (Proteintech; 1:200, Chicago, USA), mouse anti-Cdh1 (Abcam; 1:100, Cambridge, UK) or rabbit anti-Cdh1 (NBP2, Novusbio; 1:250, Cambridge, UK). Afterwards, sections were rinsed 2 times in TBS-Tx and 2 times in HEPES (10 mM) in physiological saline solution and then incubated for 3 h at RT in a solution of 2% BSA in HEPES (10 mM) with the secondary antibody: Alexa Fluor 488 anti-rabbit (CellSignaling Tech; 1:1000, Danvers, USA) and Alexa Fluor 647 anti-mouse (CellSignaling Tech; 1:1000). Finally, sections were rinsed in HEPES (10 mM), mounted onto pig skin gelatin-coated slides (0.5%) and covered with a drop of VECTASHIELD Mounting Medium with DAPI (H1200, Vector, Burlingame, USA) for nuclear staining and a drop of fluorescence mounting medium (S3023, DAKO, Barcelona, Spain) was added.