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WT MCF7 cells ( A ), MCF7 cells expressing STARD3 WT ( B – F ) or FFAT-motif deficient mutants (STARD3 FYAA ( F ), STARD3 ΔFFAT ( G )) were left untreated (left) or treated with CHIR99021 (5 µM, overnight; right). Cells were labeled with anti-STARD3 antibodies (green) ( A – G ) and in magenta with: anti-LAMP1 antibodies ( A , B ) to label <t>LE/Lys,</t> <t>anti-EEA1</t> antibodies ( C ) to label early endosomes, anti-GM130 antibodies ( D ) to label the Golgi apparatus, anti-calnexin antibodies ( E ) to label the ER or phospho-specific antibodies (pS 209 STARD3, F , G ). Nuclei were stained with Hoechst (blue). Subpanels show higher magnification images of the area outlined in white. Scale bars: 10 µm. Inset scale bars: 2 µm. .
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WT MCF7 cells ( A ), MCF7 cells expressing STARD3 WT ( B – F ) or FFAT-motif deficient mutants (STARD3 FYAA ( F ), STARD3 ΔFFAT ( G )) were left untreated (left) or treated with CHIR99021 (5 µM, overnight; right). Cells were labeled with anti-STARD3 antibodies (green) ( A – G ) and in magenta with: anti-LAMP1 antibodies ( A , B ) to label LE/Lys, anti-EEA1 antibodies ( C ) to label early endosomes, anti-GM130 antibodies ( D ) to label the Golgi apparatus, anti-calnexin antibodies ( E ) to label the ER or phospho-specific antibodies (pS 209 STARD3, F , G ). Nuclei were stained with Hoechst (blue). Subpanels show higher magnification images of the area outlined in white. Scale bars: 10 µm. Inset scale bars: 2 µm. .

Journal: The EMBO Journal

Article Title: STARD3 regulates lysosome positioning and contacts via a GSK3-controlled phosphorylation switch

doi: 10.1038/s44318-026-00705-3

Figure Lengend Snippet: WT MCF7 cells ( A ), MCF7 cells expressing STARD3 WT ( B – F ) or FFAT-motif deficient mutants (STARD3 FYAA ( F ), STARD3 ΔFFAT ( G )) were left untreated (left) or treated with CHIR99021 (5 µM, overnight; right). Cells were labeled with anti-STARD3 antibodies (green) ( A – G ) and in magenta with: anti-LAMP1 antibodies ( A , B ) to label LE/Lys, anti-EEA1 antibodies ( C ) to label early endosomes, anti-GM130 antibodies ( D ) to label the Golgi apparatus, anti-calnexin antibodies ( E ) to label the ER or phospho-specific antibodies (pS 209 STARD3, F , G ). Nuclei were stained with Hoechst (blue). Subpanels show higher magnification images of the area outlined in white. Scale bars: 10 µm. Inset scale bars: 2 µm. .

Article Snippet: The primary antibodies used were: rabbit anti-GFP (1:1000; Torrey Pine Biolabs TP401, RRID:AB_10013661), mouse anti-Lamp1 (1:50; DSHB H4A3, RRID:AB_2296838), rabbit anti-GOLGA2/GM130 (1/1000; Proteintech 11308-1-AP, RRID:AB_2115327), mouse anti-EEA1 Clone 14 (1:1000; BD Biosciences 610457, RRID:AB_397830), rabbit anti-calnexin (1:1000; Proteintech 10427-2-AP, RRID:AB_2069033) rabbit anti-STARD3 (1611; 1:2000; IGBMC), mouse anti-STARD3 (3G11; 1:100; IGBMC), rabbit anti-pS 209 STARD3 (3144; 1:200; IGBMC), and rabbit anti-STARD3NL (1545; 1:1000, IGBMC).

Techniques: Expressing, Labeling, Staining