Journal: Cell reports

Article Title: ZBTB7A is a modulator of KDM5-driven transcriptional networks in basal breast cancer

doi: 10.1016/j.celrep.2024.114991

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Goat anti-Armenian Hamster IgG (H+L) Secondary Antibody, HRP , Life Technologies , Cat# PA1–32045; RRID:AB_10985178.

Techniques: Control, Produced, Recombinant, Virus, XF Assay, ROS Assay, CRISPR, Mass Spectrometry, Knock-Out, Plasmid Preparation, Software, Combined Bisulfite Restriction Analysis Assay, Genome Wide

Journal: Cell reports

Article Title: ZBTB7A is a modulator of KDM5-driven transcriptional networks in basal breast cancer

doi: 10.1016/j.celrep.2024.114991

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Antibodies used for Immunoblotting were anti-beta-Actin (Sigma, A2228), anti-alpha Tubulin (Sigma, T5168), anti-ZBTB7A (Invitrogen, 14-3309-82), anti-KDM5A (Abcam, ab70892), anti-KDM5B (Sigma, HPA027179), anti-KDM5C (Abcam, ab34718), anti-H3K4me3 (Abcam, ab8580), anti-H3 (Active Motif, 39763), anti-RHOA (Cell Signaling Technology, 2117), anti-PKN2 (Abcam, ab87812), anti-MTA1 (Cell Signaling Technology, 5647), anti-MTA2 (Cell Signaling Technology, 15793), anti-MBD2 (Abcam, ab188474), anti-MBD3 (Cell Signaling Technology, 14540), anti-CHD3 (Cell Signaling Technology, 4241), anti-CHD4 (Cell Signaling Technology, 11912), anti-phospho-p65 (Ser536) (Cell Signaling Technology, 3033), anti-p65 (Abcam, ab32536), anti-phospho-p50 (Ser337) (Invitrogen, PA5–37658), anti-p50 (Invitrogen, MA5–15870), Goat anti-Mouse IgG Secondary HRP (Invitrogen, 62–6520), Goat anti-Rabbit IgG Secondary HRP (Invitrogen, 65–6120), and Goat anti-Armenian Hamster IgG Secondary HRP (Invitrogen, PA1–32045).

Techniques: Control, Produced, Recombinant, Virus, XF Assay, ROS Assay, CRISPR, Mass Spectrometry, Knock-Out, Plasmid Preparation, Software, Combined Bisulfite Restriction Analysis Assay, Genome Wide

Journal: Cell communication and signaling : CCS

Article Title: A non-catalytic role of IPMK is required for PLCγ1 activation in T cell receptor signaling by stabilizing the PLCγ1-Sam68 complex.

doi: 10.1186/s12964-024-01907-0

Figure Lengend Snippet: Fig. 2 IPMK depletion reduces TCR signaling through diminished activation of PLCγ1. (A) Immunoblot analysis of PLCγ1 and its phosphorylation (Y783) in lysates of naïve CD4+ T cells stimulated with anti-CD3 and anti-CD28 antibodies for 3–10 min. (B) Calcium flux of naïve CD4+ T cells activated by anti-CD3, anti-CD28, and anti-Armenian hamster IgG secondary antibodies in calcium-free HBSS. Basal time: 0–30 s; activation time: 50–300 s. ΔMFI was measured as MFIActivation time – MFIBasal time. (C) Immunoblot analysis of PKCθ and its phosphorylation (T538) in lysates of naïve CD4+ T cells stimulated with anti-CD3, anti-CD28, and anti-Armenian hamster IgG secondary antibodies for 3–10 min. (D) Quantitative real-time PCR analysis of the expression of Il2 mRNA in naïve CD4+ T cells polarized into Th0 cells by anti-IFNγ and anti-IL-4 antibodies. (E) Flow cytometry analysis of IL-2 in naïve CD4+ T cells polarized into Th0 cells for 16 h by anti-IFNγ and anti-IL-4 antibodies. All blots, kinetics, and flow cytometric plots presented are representative of at least three independent experiments. Densitometric quantitation results were normalized to control conditions. The normality of all data was analyzed using the Shapiro-Wilk test. Data are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, using the unpaired two-tailed Student’s t-test (B and E) compared to IPMKf/f or two-way ANOVA with Šídák’s multiple comparisons test (A, C, and D)

Article Snippet: Subsequently, 20 μg/mL anti-Armenian hamster IgG secondary antibody (Jackson ImmunoResearch Laboratory) was added to the RPMI 1640 medium and the cells were treated for either 3–10 min in a 37 °C heat block, after which the medium was rapidly replaced with cold PBS.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Real-time Polymerase Chain Reaction, Expressing, Flow Cytometry, Quantitation Assay, Control, Two Tailed Test