Extended Data Fig. 1. Small heat shock proteins enrich at mitochondria after heat shock in multiple human and mouse cell lines.
a, Mitochondria were isolated and compared to the cytosol and the NP-40-soluble whole cell lysate (WCL) from: HEK293T (human embryonic kidney cells), NSC-34 (motor neuron-like cells), Neuro-2A (neuronal crest-derived), SH-SY5Y (neuroblastoma cells), COS1 (African green monkey derived kidney cells), A498 (renal carcinoma cells), and healthy volunteer-derived human lymphoblasts. Cells were either left untreated or heat shocked (42 °C) for 1 h. Samples were analyzed by SDS-PAGE followed by immunoblotting using antibodies against sHSPs, Tubulin (cytosol), and VDAC1 (mitochondria). b, Mitochondria were isolated from C2C12 cells after heat shock (42 °C) for the indicated times. Samples were analyzed by SDS-PAGE followed by immunoblotting using antibodies against sHSPs or VDAC1 (loading control). Results are representative of two replicates (a,b). Unprocessed blots are available in source data.