Journal: Nature chemical biology

Article Title: A chemoproteomic portrait of the oncometabolite fumarate

doi: 10.1038/s41589-018-0217-y

Figure Lengend Snippet: Functional analyses of FH -regulated Cys residues. (a) Percentage of FH -regulated Cys residues predicted to be functional using the informatics tool Mutation Assessor (R ≥1.5, n ≥2). (b) Gene ontology analysis of FH -regulated Cys residues. (c) Correlation between genes containing FH -regulated Cys residues and genetic alterations in kidney cancer (RCC). Statistical significance was assessed using Student’s t -test (two-tailed, unpaired); ** P < 0.01. (d) SMARCC1 C520 lies in the SWIRM domain and is highly conserved. (e) SMARCC1 C520 lies at the SNF5 subunit interface. (f) SMARCC1 C520 undergoes FH -regulated changes in occupancy in HLRCC cells ( n = 3 independent experiments). (g) SMARCC1 C520E mutation limits co-immunoprecipitation with SNF5 in HEK-293 cells co-overexpressing FLAG-tagged SNF5 with Myc-tagged SMARCC1. (h) SMARCC1 S-succination can be detected in FH-deficient and FH WT HLRCC cell lines after IP of endogenous SMARCC1. (i) SNF5 demonstrates decreased co-immunoprecipitation and decreased levels in FH−/− HLRCC cells. Left: Results from SWI/SNF complex co-immunoprecipitation with BRG1 antibody. Right: Endogenous levels of SWI/SNF complex members in HLRCC cells. Representative images from two independent experiments are shown in g-i. Uncropped scans of immunoblots are provided in . (j) EZH2 inhibitors exhibit modest selectivity for FH-deficient HLRCC cells. UOK262 FH −/− or FH WT spheroids were treated with GSK126 (21 days; 1, 2, 4, 8, 16 μM) and % viability plotted relative to the vehicle treated spheroids. Data is presented as mean ± s.d.; n = 3/group. (k) EZH2 inhibitors are toxic to HLRCC spheroids. UOK262 FH −/− spheroids were treated with vehicle or EPZ6438 (14 days; 5 μM). Figure is representative of 6 replicates.

Article Snippet: SMARCC1 (11956), SNF5 (8745), BRG1 (3508), PKM1 (7067), Myc-Tag (2278), FLAG-Tag (14793) and HA-Tag (3724) antibodies were purchased from Cell Signaling Technologies.

Techniques: Functional Assay, Mutagenesis, Two Tailed Test, Immunoprecipitation, Western Blot

Journal: Ophthalmology Science

Article Title: Persistent Neutrophil Infiltration and Unique Ocular Surface Microbiome Typify Dupilumab–Associated Conjunctivitis in Patients with Atopic Dermatitis

doi: 10.1016/j.xops.2023.100340

Figure Lengend Snippet: Bacterial growth in presence of dupilumab. Acetobacter aceti and Staphylococcus aureus were cultured in different concentrations of dupilumab (2.5, 5, and 10 mg/L) and the growth was measured over a 48-hour period. A, Growth curves of A. aceti in various concentration of dupilumab. B, Area under curve (AUC) was calculated from the growth curves from 0 to 48h and data are presented in box and whisker plots showing minimum, maximum, and median values. C, Growth curves of S. aureus in various concentration of dupilumab. B, AUC was calculated from the growth curves from 0 to 48h and data are presented in box and whisker plots showing minimum, maximum, and median values. N = 8–12. One-way analysis of variance was used for statistical testing. P values are shown for respective doses vs. control. OD = optical density.

Article Snippet: Optical density at 600 nm was adjusted to ∼ 1 for Staphylococcus aureus (ATCC/ BAA-17717) in tryptic soy broth and 0.5 for Acetobacter aceti (DSMZ- 3508) in yeast peptone mannitol medium.

Techniques: Cell Culture, Concentration Assay, Whisker Assay, Control