GRP78 Quantification in MoDCs

MoDCs were seeded in a 12-well culture plate as described above (1 × 10⁶/mL/well). Western blot analyses were performed as described in Nossol et al. (2023 (link)). GRP78 was used as primary antibody (1:1000; Abcam, Germany; blocked with 5% milk) and mouse anti-β-actin as loading control (1:30,000, cell signaling, Germany). The secondary antibody was purchased with the BM Chemiluminescence Western Blotting Kit Mouse/Rabbit (Roche, Germany). Blots were analyzed on an Alpha-Ease^® FC Imaging System (Alpha Innotech, Canada). Raw intensities of protein bands were used for statistical analyses. All experiments were carried out three times. The loading control actin was utilized for normalization, and the bands with the highest raw intensity were used to normalize all other actin bands (normalization factor). Subsequently, the normalization factors were applied to the corresponding raw intensities of the investigated protein (GRP78). The normalized control values (CON-group) were used as reference in the boxplot diagrams.