Evaluating PKP2 Protein Expression Levels

PKP2 protein expression levels were evaluated in si-NC, inhibitor-NC, si-PKP2, miR-1245b-5p inhibitor, and si-PKP2+ miR-1245b-5p inhibitor cells. Transfected cells were lysed in RIPA buffer (Thermo Fisher Scientific), and GAPDH was used as a reference protein. Western blot analysis was performed as described by Wang et al. [18 (link)]. Briefly, proteins were denatured and separated using a 5% concentrate and 12% separation gel. The proteins were then transferred to polyvinylidene fluoride (PVDF; Bio-Rad, Hercules, CA, USA) membrane and incubated with rabbit anti-PKP2 (1:1000; no. #223,757; Abcam, Cambridge, UK) and anti-GAPDH (1:2500; no. #ab9485, Abcam) overnight at 4°C. Following this, the membrane was incubated with a horseradish peroxidase-conjugated goat anti-rabbit IgG secondary antibody (1:5,000; no. #ab6721; Abcam) for 1 h at room temperature. Finally, the protein bands were observed by electrochemiluminescence (EMD Millipore).