P pkd s916

The primary antibodies used were RhoA (#2117, for WB), Parkin (#2132), COX-IV (#11967), VDAC (#4661), lamin A/C (#2032), Rho-GDI (#2564), HA (#3724), PKD (#90039), P-PKD S916 (#2051), GAPDH (#2118), α-actinin (#3134) and LC3B (#3868) from Cell Signaling Technology; PINK1 from Novus Biologicals (#NB600-973); RhoA (SC-418, for IP) and ubiquitin (SC-8017) from Santa Cruz Biotechnology; miniSOG from Kerafast (#EFH004). Horseradish peroxidase (HRP)-conjugated secondary antibodies for WB, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), cycloheximide (CHX), MG-132, CID755673, Bafilomycin A1, Evans blue and triphenyltetrazolium chloride (TTC) were purchased from MilliporeSigma. Y-27632 was purchased from Cell Signaling Technology. DharmaFECT-1 and LysoTracker Blue were purchased from Thermo Fisher Scientific. C3 exoenzyme was purchased from Cytoskeletton, Inc.

GRC-2 (16-Angeloyloxy-20-acetoxy-13α-isobutanoyloxy-4β,9α-dihydroxytiglia-1,6-dien-3-one) was extracted from the aerial parts of Euphorbia grandicornis according the method described previously [20 (link)]. The purity of GRC-2 is above 95% according to its NMR spectrum and HPLC analysis. Prostratin was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for p-PKC-δ-T505, p-PKD-S744, p-PKD-S916, PKD, p-PKC-α-T638, p-cdc25c and GAPDH were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies for PKC-δ, PKC-α, p-ERK, ERK, p21, p27, cyclin B1 and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies for CDK-1 were purchased from BD Biosciences (San Diego, CA, USA). Secondary antibodies were from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, USA).