Journal: Cell

Article Title: Anti-NKG2A mAb Is a Checkpoint Inhibitor that Promotes Anti-tumor Immunity by Unleashing Both T and NK Cells

doi: 10.1016/j.cell.2018.10.014

Figure Lengend Snippet:

Article Snippet: Anti-human CD107b APC (H4B4) , Miltenyi Biotec , Cat#130-103-960; RRID: AB_2654504.

Techniques: Purification, Control, Recombinant, Selection, Staining, Software

Journal: The Journal of Biological Chemistry

Article Title: Calmodulin enhances mTORC1 signaling by preventing TSC2-Rheb binding

doi: 10.1016/j.jbc.2024.108122

Figure Lengend Snippet: Ca 2+ /CaM affects TSC2 localization without altering TSC2 phosphorylation . A , HEK293 cells were starved of serum (−FBS) for 3 h, pretreated with DMSO or BAPTA-AM (50 μM) or W-7 (30 μM) for the last 1 h (BAPTA-AM) or 5 min (W-7) and then stimulated with or without (−) carbachol (100 μM) for 15 min (+CCh). Cell lysates were analyzed by Western blotting with the indicated antibodies. The images of phospho-T389-S6K1 and S6K1 panels in DMSO and BAPTA-AM lanes are reused from Fig. 6 C . B , Quantitation of the relative intensity of phospho-T1462-TSC2 to total TSC2 of (A), in which the carbachol stimulation (DMSO) was set to 1. Graphs represent mean ± SD of three independent experiments. One-way ANOVA with Tukey’s test. C , HEK293 cells were starved of serum (−FBS) for 3 h, and then treated with carbachol (100 μM) or insulin (100 nM) for 15 min. The cells were then immunostained with anti-TSC2 and anti-LAMP2 antibodies. Merged images of TSC2 ( green ), LAMP2 ( magenta ) and nuclei staining with Hoechst 33342 ( blue ) are also shown. Scale bar, 10 μm. D , colocalizations of TSC2 with LAMP2 were quantified, and Pearson’s correlation coefficient is shown. Data from four independent experiments are shown in blue triangles and were used for statistical analysis. Bars represent averages. The values of Pearson’s correlation coefficient of five individual images in each experiment are also shown in gray circles . One-way ANOVA with Tukey’s test, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: For immunofluorescence, TSC2 (D93F12, Cell Signaling Technology #4308) and LAMP2 (H4B4, Proteintech 65053-1-Ig) were used as primary antibodies.

Techniques: Phospho-proteomics, Western Blot, Quantitation Assay, Staining