Extended Data Fig. 7: FSP1 preferentially localizes to perinuclear lysosomes in LN metastatic lines.
From: Lymph node environment drives FSP1 targetability in metastasizing melanoma
a, Left, representative confocal microscopy of FSP1-OFP (Green) with lipid droplets (LipidSpot: Red), mitochondria (Mitoview: Magenta), and nuclei (Hoechst; Blue) in LN7 and LN8 lines. Right, histogram of the fluorescence intensity profile across the arrow. b,c, Immunoblots of Golgi- (b) and ER-enriched (c) fractions. RCAS1 and PDI3A were used as Golgi and ER markers, respectively and γ-tubulin served as a whole-cell extract control. ER extract (b), n = 4; Golgi extract (c), n = 4 independent experiments. d, Representative confocal microscopy of FSP1-OFP (Green) and LAMP1 (Magenta) ± IMP-1088 (0.1 μM) in B16-F0, LN7, LN8, and LN9 lines for 24 h. e, Representative confocal microscopy of FSP1-OFP (Green) and Lysotracker (Magenta) in B16-F0, LN7, LN8, and LN9 lines. f, Representative confocal microscopy of FSP1-OFP (Green) and Lysotracker (Magenta) localization in LN8 1194BR ± IMP-1088 (0.1 μM) under 1% O2 for 24 h. g, Representative confocal microscopy of FSP1-OFP WT and FSP1 OFP G2A (Green) localization with LAMP1 (Magenta) in LN9 1315BL. h, Representative confocal microscopy of FSP1-OFP (Green) and Lysotracker (Magenta) in SK-MEL5, MeWo and A-375 cells. i, FSP1 subcellular localization in LN metastatic lines. The diagram was created using BioRender. 3 independent experiments for a, d-h. Scale bar 10 μm (a, d, e, f, h) and 5 μm (g).
