Alexa fluor 594 goat anti rabbit secondary antibody

Cell samples were fixed in 4% paraformaldehyde in PBS for 15 min at room temperature. Tissue samples were fixed in 4% paraformaldehyde in PBS overnight at room temperature. The tissue samples were embedded, frozen, and sliced. Both the cell and tissue samples were then washed, permeabilized, and blocked. We used the following antibodies to investigate protein expression in these samples: anti-Piezo1 (Abcam, ab128245, 1:100), anti-Ki67 (CST, #9449, 1:400), anti-F-actin (Abcam, ab130935, 1:500), anti-α-SMA (CST, #48938, 1:200), Alexa Fluor 594 goat anti-rabbit secondary antibody (Jackson lab, 125369, 1:200), and Alexa Fluor 488 goat anti-mouse secondary antibody (Jackson lab, 133384, 1:200). Images were captured using a confocal microscope (Zeiss, Germany) or a Nikon Eclipse E800 microscope (Nikon, Melville, NY, USA).

Cell samples were fixed with 4% PFA for 15 min at RT. Tissues were fixed with 4% PFA overnight at RT. Tissue samples were embedded in paraffin and sliced into 5 mm sections. Samples were incubated with following antibodies: Piezo1 (ab128245, 1:100, Abcam), α-SMA (ab7817, 1:100, Abcam), CCL24 (22306-1-AP, 1:200, Proteintech), CCR3 (ab32512, 1:100, Abcam), Wnt2 (A5864, 1:200, Abclonal, Wuhan, China), Wnt11 (ab31962, 1:200, Abcam), β-catenin(#8480, 1:100, CST), Alexa Fluor 594 goat anti-rabbit secondary antibody (Jackson lab, 125369, 1:200) and Alexa Fluor 488 goat anti-mouse secondary antibody (Jackson lab, 133384, 1:200). Images were visualized using a Nikon Eclipse E800 microscope (Nikon, Melville, NY, USA).

Samples were fixed with 4% paraformaldehyde overnight, then embedded in paraffin. 5 mm sections were processed and stained with hematoxylin and eosin (H&E) and Masson’s Trichrome Stain Kit following protocol (Solarbio, Beijing, China).
For immunohistochemistry and immunofluorescence, 5 mm sections were incubated with primary antibodies against Piezo1 (ab128245, 1:1000, Abcam), α-SMA (ab7817, 1:200, Abcam), Collagen I (ab6308, 1:200, Abcam), GFP (ab290, 1:500, Abcam), Arg1 (16001-1-AP, 1:400, Proteintech), OAT (A6235, 1:100, ABclonal Technology), PYCR1 (13108-1-AP, 1:200, Proteintech), PRODH (22980-1-AP, 1:400, Proteintech) overnight at 4 °C. Sections were then incubated with Alexa Fluor 594 goat anti-rabbit secondary antibody (125369, 1:200, Jackson lab), and Alexa Fluor 488 goat anti-mouse secondary antibody (133384, 1:200, Jackson lab) for immunofluorescence or an HRP-conjugated goat anti-rabbit secondary antibody (138729, 1:500, Jackson lab) for immunohistochemistry. Images were captured using a Nikon Eclipse E800 microscope (Nikon, Melville, NY, USA).