Journal: Cell Reports

Article Title: PIM kinases regulate early human Th17 cell differentiation

doi: 10.1016/j.celrep.2023.113469

Figure Lengend Snippet: PIMs regulate Th1/Th17 signaling axis during early Th cell differentiation (A) Based on bulk RNA-seq data, STAT1 was predicted as a positive upstream regulator of the differentially expressed genes at 6 h upon PIM TKD using the IPA “upstream regulator” prediction tool ( Z score < −2, which indicates predicted upstream regulators; gray arrows represent effects not predicted. (B) The downregulation of STAT1 RNA levels was confirmed at 24 h and 72 h in PIM TKD Th17 cells by qRT-PCR. (C and D) The RNA expression of Th1-related factors TBX21 (C) and STAT4 (D) was validated in PIM TKD at 6 h and 24 h of Th17 cell differentiation by qRT-PCR. FC normalized to the Scr control was plotted for four biological replicates. Boxplots represent median and interquartile range, and whiskers extend to maximum and minimum values (B–D). Statistical significance is calculated from four biological replicates using two-tailed Student’s t tests ( ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (E and F) scRNA-seq was performed at 6 h in PIM TKD and control Th17 cells using the single-cell fixed RNA profiling. Cells double-negative for PIM – /TBX21 but RORA positive (E) and single cells double-positive for PIM/TBX21 but RORA -negative (F), were colored in blue and projected into a two-dimensional map using UMAP. (G) Differential expression analysis was performed between Scr (control) and PIM – in PIM TKD samples at 6 h of differentiation (FDR of <0.05, log2FC of >0.24) for one biological replicate. Scaled scRNA-seq dot plot depicting the differentially expressed genes of interest on the x-axis. The color scale represents the average expression of a given gene in the cluster, and the size of the dot represents the percent of cells that express a given gene. (H) Violin plots showing the expression of CD82 and STAT1 in Scr and enriched PIM – cells of PIM TKD samples at 6 h. See also Figures S4 and and Table S2 .

Article Snippet: rabbit monoclonal anti-human STAT4 , Cell Signaling Tech , Cat# 2653; RRID: AB_2255156.

Techniques: Cell Differentiation, RNA Sequencing, Quantitative RT-PCR, RNA Expression, Control, Two Tailed Test, Quantitative Proteomics, Expressing

Journal: Cell Reports

Article Title: PIM kinases regulate early human Th17 cell differentiation

doi: 10.1016/j.celrep.2023.113469

Figure Lengend Snippet:

Article Snippet: rabbit monoclonal anti-human STAT4 , Cell Signaling Tech , Cat# 2653; RRID: AB_2255156.

Techniques: Recombinant, Sequencing, Purification, Isolation, Enzyme-linked Immunosorbent Assay, DC Protein Assay, Gene Expression, Reverse Transcription, Staining, CRISPR, Plasmid Preparation, Software

Journal: The Journal of pharmacology and experimental therapeutics

Article Title: Generation and Characterization of Mirikizumab, a Humanized Monoclonal Antibody Targeting the p19 Subunit of IL-23.

doi: 10.1124/jpet.122.001512

Figure Lengend Snippet: Fig. 2. Miri (LY3074828) specifically inhibits IL-23 but not IL-12 signaling in vitro. Ratio of pSTAT4:STAT4 in Kit225 (A) or TALL-104 (B) cells following stimulation with IL-12 in the presence of the miri/IL-23 complex.

Article Snippet: Rabbit anti-human phospho-STAT4, mouse anti-human STAT4, and rabbit anti-human b-actin monoclonal antibodies (all from Cell Signaling) were used at 1:1000 dilution.

Techniques: In Vitro

Journal: Immunity, Inflammation and Disease

Article Title: Interleukin‐23 receptor defines T helper 1‐like regulatory T cells in oral squamous cell carcinoma

doi: 10.1002/iid3.746

Figure Lengend Snippet: Flow cytometry antibodies

Article Snippet: PE anti‐mouse/human Phospho‐STAT4 (Tyr693) , MA5‐37332 , Stat4Y693‐F6 , ThermoFisher.

Techniques: Flow Cytometry