Rhod dextran

Carbonyl cyanide 3-chlorophenylhydrazone (CCCP), ethylene glycol tetra acetic acid (EGTA), hydrogen peroxide solution (H₂O₂), d-mannitol, and lipopolysaccharides were purchased from Sigma-Aldrich, USA. Bafilomycin A1 and GPN were from Santa Cruz Biotechnology, while BAPTA-AM, cyclosporin A, FK506, ionomycin, nicotinic acid adenine dinucleotide phosphate (NAADP), trans-Ned-19 (Ned-19), and U18666A were from Tocris Biosciences, USA. Thapsigargin (TG) was bought from Almone Labs, USA, while LysoTracker Red DND-99 Dye and Rhod dextran were from Invitrogen, USA. Antibodies used for immunoblotting and immunostaining were as follows: anti-mouse lysosome-associated membrane protein-1 (LAMP-1; sc-20011, Santa Cruz Biotechnology, USA), anti-rabbit cathepsin D (2284S, Cell Signaling, USA), anti-rabbit caspase-1 (2225S, Cell Signaling, USA), anti-rabbit TFEB (37785S, Cell Signaling, USA), anti-rabbit histone H3 (D1H2) (4499S, Cell Signaling, USA), anti-rabbit Integrin β1 (4706S, Cell Signaling, USA), anti-rabbit GAPDH (2118S, Cell Signaling, USA), and anti-rabbit α/β-tubulin (2148S, Cell Signaling, USA).

Calcium concentrations were quantified as described
^{22 (link)} with low-affinity Rhod-dextran (
Kd=551 ± 107μM) (Invitrogen) in conjunction with the calcium-insensitive Alexa-Fluor 488 dextran (Invitrogen) at concentrations of 0.25mg/ml and 0.1mg/ml, respectively. Dextrans were loaded for 12hr, followed by a 12hr chase.