67568 2 ig  (Proteintech)

Journal: Cell reports

Article Title: STAT-independent functions of Janus kinases 1 and 2 are obligatory for the postnatal development of mammary epithelial ducts

doi: 10.1016/j.celrep.2025.116703

Figure Lengend Snippet: (A) Schematic of the knockout of STAT1, STAT3, STAT5a, and STAT5b in canonical JAK/STAT signaling cascades of the mammary gland. (B) Carmine alum-stained mammary gland wholemounts of nulliparous female mice with a targeted deletion of four Stat genes in the mammary epithelium MMTV-Cre Stat3/5 fl/fl Stat1 −/− ) and a STAT1 single-knockout littermate control; bars, 1 mm. (C) Immunoblot analysis of the seven known mammalian STAT proteins in mammary epithelial cells (MECs) from STAT1/3/5a/5b quadruple-knockout females ( N = 4) in comparison to age-matched STAT3/5a/5b triple-knockout mice ( N = 2). Other controls: C1 and C2, positive controls for active STAT3 and STAT5, mammary gland tissues from wild-type mice on day 1 of involution and lactation, respectively; C3, spleen as a positive control for STAT4; C4, wild-type MECs as a negative control for STAT4; C5 and C6 interferon-treated and untreated MECs as positive and negative controls for STAT2; C7 and C8, IL-4-treated and untreated wild-type MECs as positive and negative controls for active STAT6. GAPDH and ACTB served as loading controls.

Article Snippet: Rabbit monoclonal, STAT4 , Cell Signaling , Cat#2653; RRID:AB_ 2255156.

Techniques: Knock-Out, Staining, Control, Western Blot, Quadruple Knockout, Comparison, Triple Knockout, Positive Control, Negative Control

Journal: Cell reports

Article Title: STAT-independent functions of Janus kinases 1 and 2 are obligatory for the postnatal development of mammary epithelial ducts

doi: 10.1016/j.celrep.2025.116703

Figure Lengend Snippet: (A and B) Immunoblot analysis of tyrosine-phosphorylated JAK1 and JAK2 and expression of selected STAT proteins in mammary epithelial cells from quadruple STAT1/3/5a/5b conditional knockout mice and wild-type controls that were treated with either oncostatin M (OSM) alone and human growth hormone (hGH) alone (A) or a combination of both (B); 2 biological repeats of experimental and control animals. GAPDH served as a loading control. (C) Immunoblot analysis to assess the activation of STAT6 and tyrosine phosphorylation of JAK1 in STAT1/3/5a/5b-deficient quadruple-knockout cells and controls following stimulation with IL-4. GAPDH was used as a loading control. PC, splenocytes as positive controls for the validated absence of STAT2 and STAT4 in the quadruple-knockout epithelial cells. (D) Summary of canonical and noncanonical signaling mechanisms by which JAK2, in cooperation with JAK1, drives the postnatal development of the mammary gland.

Article Snippet: Rabbit monoclonal, STAT4 , Cell Signaling , Cat#2653; RRID:AB_ 2255156.

Techniques: Western Blot, Expressing, Knock-Out, Control, Activation Assay, Phospho-proteomics, Quadruple Knockout

Journal: Human Cell

Article Title: Ethambutol induces optic neuropathy through SDHB-mediated ferroptosis in retinal ganglion cells via Smad4 pathway

doi: 10.1007/s13577-025-01342-4

Figure Lengend Snippet: EMB modulates SDHB transcriptional activity via Smad4 in RGC-5 cells. A WB analyses were conducted to assess the NFκB, Smad2/3, Smad4, STAT4 levels, and their phosphorylation levels in RGC-5 cells treated with 2 mM EMB. N = 3. * p < 0.05, ** p < 0.01. B Utilizing the JASPAR database, a prediction was made regarding the binding of Smad4 to the SDHB promoter. C Detection of SLC7A11 and GPX4 protein expression in Smad4-overexpressing RGC-5 cells by WB. N = 3. * p < 0.05. D WB assay was conducted to verify the Smad4 overexpression in RGC-5 cells. N = 3. * p < 0.05. E , F RT-qPCR and WB assays were performed to quantify SDHB mRNA ( E ) and protein ( F ) levels in Smad4-overexpressing RGC-5 cells. N = 3. * p < 0.05. G Dual-luciferase reporter assays were employed to demonstrate EMB’s regulation of SDHB promoter activity via Smad4. SDHB-Mut: mutating the Smad4 binding sites on SDHB promoters. N = 3. * p < 0.05. H CHIP assay to detect DNA enriched fragments of SDHB promoter was conducted using Smad4-specific antibodies in EMB-treated RGC-5 cells. N = 3. ** p < 0.01. I WB analysis of Smad4 knockdown efficiency in RGC-5 cells. N = 3. * p < 0.05. J In SDHB-overexpressing RGC-5 cells with Smad4 knockdown, the protein expression levels of SDHB, SLC7A11, and GPX4 were detected by WB. Data shown are representative of three independent experimental replicates

Article Snippet: STAT4 , Proteintech , 67568-2-Ig.

Techniques: Activity Assay, Phospho-proteomics, Binding Assay, Expressing, Over Expression, Quantitative RT-PCR, Luciferase, Knockdown