Journal: Nature

Article Title: CRISPR screens decode cancer cell pathways that trigger γδ T cell detection

doi: 10.1038/s41586-023-06482-x

Figure Lengend Snippet: (a) Surface BTN3A MFI in Daudi-Cas9 KOs cultured in different pyruvate concentrations for 3 days in RPMI (no glucose, no pyruvate). Normalized to cells grown without pyruvate (0 mM). (b-d, f) Surface BTN3A MFI in Daudi-Cas9 cells treated for 72 hours with (b) an mTOR inhibitor (rapamycin), an ISR inhibitor (ISRIB), ISR agonists (guanabenz, Sal003, salubrinal, raphin1, sephin1), and DMSO (vehicle) (KO cells); (c) metformin (WT cells); (d) A-769662 compared to equivalent amounts of DMSO (vehicle) (WT cells); or (f) the shown compounds (KO cells). (e) Surface BTN3A MFI in WT Daudi-Cas9 cells co-treated with AICAR and increasing amounts of Compound C (AMPK inhibitor) or DMSO (vehicle). (a) n=4 per condition (n=3, TIMMDC1 (#2) at 0 mM), data combined from two independent experiments, each individually normalized. (b) n=6 per condition (except n=5 for AAVS1 (#5) with guanabenz and for PPAT (#1) with salubrinal) , data combined from two independent experiments, each individually normalized to DMSO (vehicle)-treated cells. (c) n=8 per condition, data combined from two independent experiments. One-way ANOVA comparison to cells that received no treatment with Dunnett’s multiple comparisons test. (d) n=3 per condition, representative data from one of two independent experiments. (e) n=3 per conditions, representative data from one of two independent experiments. Two-tailed unpaired Student’s t test with Bonferroni correction. (f) n=3 per condition (n=2 for AMPKα1 (#1) treated with DMSO), representative data from one of two independent experiments. One-way ANOVA comparison to AAVS1 (#5) KO cells with Dunnett’s multiple comparisons test. (a-f) Mean ± SD. p<0.0001 (****), p<0.001 (***), p<0.01 (**), p<0.05 (*), p>0.05 (N.S.).

Article Snippet: Cells received 10 μL of DMSO (vehicle) or one of the following compounds at a final concentration of 10 μM: sephin1 (APExBIO), ISRIB (MedChemExpress), guanabenz acetate (MedChemExpress), Sal003 (MedChemExpress), salubrinal (MedChemExpress), raphin1 acetate (MedChemExpress), and rapamycin (MilliporeSigma).

Techniques: Expressing, Cell Culture, Comparison, Two Tailed Test

Journal: Cell Stem Cell

Article Title: Androgen Signaling Regulates SARS-CoV-2 Receptor Levels and Is Associated with Severe COVID-19 Symptoms in Men

doi: 10.1016/j.stem.2020.11.009

Figure Lengend Snippet:

Article Snippet: Guanabenz acetate , Selleck Chem (FDA-Approved Library) , N/A.

Techniques: Control, Virus, Recombinant, Purification, SYBR Green Assay, Staining, Gene Knockout, Transfection, RNA Sequencing, Plasmid Preparation, Expressing, Software, Generated

Journal: Experimental neurology

Article Title: Guanabenz promotes neuronal survival via enhancement of ATF4 and parkin expression in models of Parkinson disease.

doi: 10.1016/j.expneurol.2018.01.015

Figure Lengend Snippet: Fig. 6. ATF4 and parkin are necessary for guanabenz- mediated neuroprotection against 6-OHDA. (A–C) Differentiated PC12 cells were infected with lentivirus carrying shRNA against ATF4 (shATF4, in A, or Parkin (shParkin, in C), or a control shRNA (shCTRL, a mutated version of the shATF4 sequence) for 3 days. In B, cells were treated with 20 nM ISRIB (I) at the same time as GA, where indicated. Whole cell lysates were analyzed by Western blotting with the indicated antibodies; for A and B, cells were treated with 100 μM 6-OHDA to induce robust ATF4 expres- sion. In A, duplicate wells of cells infected with shATF4 were analyzed. For survival experiments, cells were pretreated with 2.5 μM GA (+/−20 nM ISRIB in B) for 4 h, then treated with 100 μM 6-OHDA for 24 h. Viable nuclei were counted. Data are mean +/−SEM from 5 independent experiments; each condition was performed in triplicate in each experiment. (D) Cultured ventral midbrain neurons were infected with lentivirus carrying shRNA against Parkin or a scram- bled control for 4 days. Then cultures were pretreated with 2.5 μM GA for 4 h, treated with 50 μM 6-OHDA for 24 h, and fixed and immunostained for GFP and TH. The number of TH/GFP+ cells on each coverslip was counted. Data are mean +/−SEM from 4 in- dependent experiments (except for GA alone with ei- ther shRNA construct, which was performed in 2 ex- periments); each condition was performed either in duplicate or triplicate. **p < 0.01, ***p < 0.001 by ANOVA with Tukey's post-hoc test.

Article Snippet: Stock solutions of 6-hydroxydopamine (6OHDA; Tocris), guanabenz acetate salt, clonidine, efaroxan (Sigma) were prepared in ddH2O; camptothecin (CPT; Tocris) was prepared in DMSO.

Techniques: Infection, shRNA, Control, Sequencing, Western Blot, Cell Culture, Construct