Journal: bioRxiv

Article Title: Mechanoresilience of lysosomes conferred by TMEM63A

doi: 10.64898/2025.12.18.695245

Figure Lengend Snippet: A) LysoFlipper FLIM in WT RAW264.7 macrophages incubated with 1 mM LLOMe for 5 or 15 min, or 250 µM GPN for 15 min. Each dot represents one field of 5-10 cells. N = 3. B) qPCR for TMEM63A, B, and C in RAW264.7, BMDM, or MEF. n = 3. C-D) TMEM63A-OFP (magenta) and LAMP1-GFP (green) in WT RAW264.7 cells before and after challenge with IgG-coated microspheres. E-F) 10 kDa dextran or SRG pulsed for 16 h and chased for 1 h in WT and TMEM63A KO BMDM. G) Lysosomal pH in WT and TMEM63A KO BMDM determined using ratiometric measurements of 10 kDa Oregon Green dextran pulsed for 16 h and chased for 1 h. Each data point represents a field containing >5 cells. n = 3. H) LAMP1, cathepsin C expression and processing by WB. I) DQ-BSA signal of WT and TMEM63A KO BMDM after pulse (1 h) and chase (1 h). All data points represent a field containing 4-8 cells. n = 3. J-K) Phagocytosis in WT and TMEM63A KO BMDM, quantified in K. Each data point represents 3-5 fields containing 15-25 cells. n = 3, scale bars 10 µm.

Article Snippet: Primary antibodies against LAMP1 (DSHB, 1D4B), Cathepsin C (Santa Cruz, sc-74590) and GAPDH (Santa Cruz, 365062) were used at 1:1000 (v/v) for western blotting.

Techniques: Incubation, Expressing