Journal: Advanced Science
Article Title: TBC1D22B Regulates ER‐to‐Golgi Trafficking via RAB1B Inactivation and Promotes Oncogenic Programs in Breast Cancer
doi: 10.1002/advs.202502269
Figure Lengend Snippet: Overexpression of TBC1D22B, but not of TBC1D22A, induces tumor spheroids overgrowth, in a GAP‐dependent manner. BT549 cells stably expressing the empty vector (EV), HA‐tagged TBC1D22B, RQ or TBC1D22A were grown in matrigel for 7 days to form tumor cell spheroids. Spheroids were stained with Phalloidin and DAPI shown respectively in magenta and grey in the merged representative images (left). Images are confocal Z‐stack projections (8 optical sections/image) of spheroids acquired with a 40x objective, zoom 1.5x. Bar, 50 µm. Quantification of the spheroids area, shown in the middle panel, is from 3 independent experiments in which 45–71 spheroids/condition were measured. Mean ± SD are also shown in the plots. Statistical analysis was performed with the t‐test, **** p < 0.0001. The Western blots on the right were IB with the indicated antibodies (vinculin, loading control) and show the expression levels of TBC1D22B, RQ and TBC1D22A. B) CAL120 cells stably expressing the empty vector (EV), the HA‐tagged TBC1D22B, or RQ were grown and stained as in A. Images are confocal Z‐stack projections (8 optical sections/image) of representative spheroids acquired with a 20x objective, zoom 2. Bar, 50 µm. In the middle panel the area of 45 spheroids/condition from 3 independent experiments is shown. Mean ± SD are also shown in the plots. Statistical analysis was performed with the t‐test, p values: ***, p < 0.001, **** p < 0.0001. The Western blot on the right shows the expression levels of TBC1D22B and RQ.
Article Snippet: CAL120 cells, from DSMZ, were grown in DMEM (ECM0749L, Euroclone) supplemented with 10% fetal bovine serum (ECS1800D, Euroclone), 1% L‐glutamine (ECB3000D, Euroclone) and 1% penicillin‐streptomycin (ECB3001D, Euroclone).
Techniques: Over Expression, Stable Transfection, Expressing, Plasmid Preparation, Staining, Western Blot, Control
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