Sc 166729

To detect the dex-induced YFP-tagged proteins, the blot was probed with anti-GFP (Clontech, 632381, 1:5,000) primary antibodies. To detect HA-tagged proteins, the blot was probed with anti-HA HRP-conjugated antibody (Cell Signaling Tech, 2999, 1:3,000). To detect endogenous proteins, the blot was probed with anti-ARF2 primary antibody (PhytoAB, PHY2435A, 1:2,000), anti-CH1 primary antibody (PhytoAB, PHY1909S, 1:2,000), anti-RBOHD primary antibody (Agrisera, AS15 2962, 1:2,000), anti-ICS1 primary antibody (Agrisera, AS16 4107, 1:2,000) or anti-β-tubulin primary antibody (Santa Cruz Biotech, sc-166729, 1:2,000). For secondary antibodies, anti-rabbit-HRP antibody (Cell Signaling Tech, 7074, 1:3,000) or anti-mouse-HRP antibody (Abcam, Ab97040, 1:10,000) were used.

Western blot and IHC staining followed established protocols (Yan et al. 2021 (link)). IHC scoring utilized image pro plus software, analyzing images to determine IOD/Area or IOD Sum values. Antibodies used included LOXL1 (1:1000, PA5-79,609, Invitrogen), β-tubulin (1:1000, sc-166729, Santa Cruz) for western blotting, and LOXL1 (1:500), CD8 (ZA-0508, ZSGB-Bio) for IHC.

Assays were performed as described previously^{19 (link)}. Beta-tubulin (sc-166729, 1:500, Santa Cruz Technology) and LaminB1 (sc-374051, 1:500, Santa Cruz Technology) were used as the cytosolic and nuclear loading control, respectively.