Mouse anti stat3

Thermo Fisher Scientific supplied mouse anti-E-cadherin, mouse anti-vimentin, mouse anti-catenin, mouse anti-galectin, mouse anti-PD-L1, mouse anti-TGF, mouse anti-NF-κB antibodies, mouse anti-STAT3, and Hoechst (Waltham, MA, USA). eBio-science provided mouse anti-CD163-PerCP, mouse anti-CD206-PerCP, mouse anti-Ki-67-APC, mouse anti-CD68-FITC, and mouse anti-IL-10-FITC (San Diego, CA, USA). BD Cytofix/Cytoperm Fixation/Permeabilization Solution Kit with GolgiPlug™(No. BDB555028, San Diego, CA, USA). PeproTech delivered recombinant mouse IL-4, IFN-, and anti-IL-10 receptor (abIL-10R) inhibitors (Rocky Hill, NJ, USA).
mouse anti-STAT3 (Invitrogen, cat number MA1-13042), rabbit anti-NF-κB (Life technologies, cat number 510500), mouse anti-PD-L1/CD274 (Proteintech, cat number 66248-1-Ig), and mouse anti-TGF (Invitrogen, cat number MA5-15065) were used for immunoblotting, and so were secondary antibodies goat anti-mouse and goat anti-rabbit Alexa® Fluor 555 (Thermo Fisher Scientific, Waltham). For immunoblotting, rabbit anti-actin (LI-COR Biosciences, Lincoln, Nebraska, USA) was used. The anti-mouse PD-L1 checkpoint blocker was purchased from InVivoPlus (B7-H1, Bio cell, USA).

Mouse anti-E-cadherin, mouse anti-vimentin, mouse anti-STAT3, mouse anti-NF-κB antibodies, and Hoechst were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Cer (from porcine brain) was purchased from Avanti Polar Lipids (Alabama, USA), PA was purchased from Sigma-Aldrich (Darmstadt, Germany), and anti-mouse CD163- PerCP, anti-mouse KI-67-APC, anti-mouse CD68-FITC, and anti-mouse IL-10-FITC were obtained from eBioscience (San Diego, CA, USA). Recombinant mouse IL-4, IFN-δ, and anti-IL-10R inhibitor were purchased from PeproTech (Rocky Hill, NJ, USA).
For immunofluorescence labeling, purified rabbit anti-Vimentin, rabbit anti NF-κB, rabbit anti-E-cadherin, and mouse anti-STAT3, and the secondary antibodies goat anti-mouse and goat anti-rabbit Alexa® Fluor 555 (Thermo Fisher Scientific) were used. Rabbit anti-β-actin (LI-COR Biosciences, Lincoln, Nebraska, USA) were used for immunoblotting.

Western blot was performed as reported in Fullone et al. 2022 [5 (link)]. The following primary antibodies were used at a dilution of 1:1000 in the blocking solution: rabbit anti-ERK (Santa Cruz Biotechnology, Dallas, TX, USA, sc-153), mouse anti-p-ERK (Cell Signaling Technology, Danvers, MA, USA, 9106S), monoclonal anti polyhistidine peroxidase (Sigma-Aldrich, MERCK, Darmstadt, Germany A7058), rabbit anti-MRAP2 polyclonal antibody (Invitrogen-Thermo Fisher Scientific, Waltham, MA, USA, PA5-113283), mouse anti-PKR2 polyclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA, sc-365696), mouse anti-STAT3, and rabbit anti-pSTAT3 (Tyr705) (1:1000, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). After extensive washing with T-TBS, membranes were incubated with the appropriate IgG HRP-linked secondary antibody for 1 h at room temperature. The immunoreactive signals were visualized using an enhanced chemiluminescence system.