Journal: Journal of Translational Medicine

Article Title: IL-27 attenuates airway inflammation in a mouse asthma model via the STAT1 and GADD45γ/p38 MAPK pathways

doi: 10.1186/s12967-016-1039-x

Figure Lengend Snippet: The STAT1 and GADD45γ/p38 MAPK pathways remain impairment in the IL-27 treatment groups. a Protocol of OVA-induced allergic asthma and IL-27 administration. b The mRNA expression of STAT1, GADD45-γ and p38 MAPK by qPCR. c The protein expression of STAT1, GADD45γ and p38 MAPK by Western blot. In IL-27 treated group, p-STAT1, p-p38MAPK were still impaired compared with PBS group, while GADD45γ had no markedly difference. d Densitometer measurements of pSTAT1/GAPDH, GADD45γ/GAPDH and p38 MAPK/GAPDH. IL-27 treatment had no improvement in the already impaired signal pathway.(GAPDH expression was acted as internal reference.) The columns and error bars represent the mean and SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001. Similar results were obtained in at least three independent experiments

Article Snippet: The following antibodies were also used: anti-STAT1 antibody (SC592; Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-Py-STAT1 antibody (catalog no. 9171; Cell Signaling, Boston, MA, USA), anti-GADD45γ antibody (ab196774,Abcam,Cambridge,UK), anti-p38 MAPK antibody (CST9212, Cell Signaling, Boston, MA, USA), and anti-phospho-p38 MAPK antibody (CST9215, Cell Signaling, Boston, MA, USA).

Techniques: Expressing, Western Blot

Journal: Journal of Translational Medicine

Article Title: IL-27 attenuates airway inflammation in a mouse asthma model via the STAT1 and GADD45γ/p38 MAPK pathways

doi: 10.1186/s12967-016-1039-x

Figure Lengend Snippet: The STAT1 and GADD45γ/p38 MAPK pathways are up-regulated in the IL-27 prevention groups. a Protocol of OVA-induced allergic asthma and IL-27 administration. b The mRNA expression of STAT1, GADD45-γ and p38 MAPK by qPCR. It showed IL-27 preventative treatment reverse the impaired STAT1 and strengthen GADD45-γ expression as well. c The protein expression of STAT1, GADD45-γ and p38 MAPK by Western blot. It shows IL-27 preventative treatment reverse the impaired p-STAT1 and strengthen GADD45-γ/p38 MAPK protein expression as well. d Densitometer measurements of pSTAT1/GAPDH, GADD45-γ/GAPDH and p38 MAPK/GAPDH. GAPDH expression was acted as internal reference. The columns and error bars represent the mean and SEM (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001. Similar results were obtained in at least three independent experiments

Article Snippet: The following antibodies were also used: anti-STAT1 antibody (SC592; Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-Py-STAT1 antibody (catalog no. 9171; Cell Signaling, Boston, MA, USA), anti-GADD45γ antibody (ab196774,Abcam,Cambridge,UK), anti-p38 MAPK antibody (CST9212, Cell Signaling, Boston, MA, USA), and anti-phospho-p38 MAPK antibody (CST9215, Cell Signaling, Boston, MA, USA).

Techniques: Expressing, Western Blot

Journal: FEBS letters

Article Title: Inhibition of matrix metalloproteinase-9 by interferons and TGF-beta1 through distinct signalings accounts for reduced monocyte invasiveness.

doi: 10.1016/j.febslet.2005.09.012

Figure Lengend Snippet: Fig. 4. Western blot analysis of STAT1, pY-STAT1 and IRF-1 proteins in monocytes treated with IFNs, TGF-b1 and AG490. Monocytes (1 · 106) were cultured in the absence or presence of IFNs (1000 U/mL) or TGF-b1 (10 ng/mL) for 24 h. Cells were pretreated with AG490 (5 lM) for 30 min before cytokine stimulation for 24 h. (A) Western blot analyses were performed as described in Section 2. Whole cell lysates were separated by 12% SDS– PAGE. (B) MMP-9 production in conditioned media of day-1 monocytes was assayed by ELISA. Data are calculated as percent of control (no stimulation or AG490 stimulation alone) and represent the mean of three separate experiments ± S.D.

Article Snippet: Membranes were hybridized with Abs for phosphorylated-STAT1 (pY-STAT1), STAT1, IRF-1 and actin as described [30] using chemiluminescence technique developed by Santa Cruz.

Techniques: Western Blot, Cell Culture, SDS Page, Enzyme-linked Immunosorbent Assay, 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) Immunoblot analysis of the expression and activation of selected STAT proteins in mammary epithelial cells from STAT3 and STAT5a/b conditional triple-knockout mice (MMTV-Cre Stat3/5 fl/fl ; N = 4) and age-matched Stat3/5 fl/wt controls ( N = 2 and 4, respectively). E-cadherin (CDH1) and beta-actin (ACTB) were used as loading controls. PC, wild-type control epithelial cells treated with IL-4 served as a positive control for tyrosine-phosphorylated STAT6 (pY-STAT6). (B) Immunohistochemistry of tyrosine-phosphorylated STAT1 (pY-STAT1) on histologic sections of mammary glands from postpartum STAT3/5 triple-knockout and control females; bars, 20 μm. (C and D) Immunoblot analysis of active and total levels of STAT1 before and after the retroviralbased expression of Cre recombinase in three immortalized mammary epithelial cell lines (C) and three mouse embryonic fibroblast lines (D). Wild-type mammary epithelial cells and interferon gamma-treated wild-type mouse fibroblasts served as positive controls (C) for STAT5a and active STAT1, respectively. The densitometry results of the STAT proteins in (C) from 3 technical repeats of the 3 biological repeats are shown in . (E) Immunohistochemistry of active STAT1 (pY-STAT1) in alveolar cells of mammary glands of a postpartum WAP-Cre Stat3/5 fl/fl female and an age-matched control; bars, 20 μm.

Article Snippet: Rabbit polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

Techniques: Western Blot, Expressing, Activation Assay, Triple Knockout, Control, Positive Control, Immunohistochemistry

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 polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

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) Immunoblot analysis of STAT1 and JAK1 expression and activation in response to the pharmacological inhibition of JAK1 using 1 μM itacitinib in three mammary epithelial cell lines co-deficient in STAT3, STAT5a, and STAT5b. Beta-actin (ACTB) was used as a loading control. The densitometry results of immunoblots from 2 technical repeats of the 3 biological repeats are shown in . (B) Schematic of a quadruple knockout of STAT3, STAT5a, and STAT5b along with JAK1 to genetically ablate the compensatory activation of STAT1 in the STAT3/5a/5b triple-knockout mammary epithelium and to determine the STAT-independent contribution of JAK2 to mammary gland development. (C) Carmine alum-stained mammary gland whole-mounts of 4- and 6-week-old nulliparous females that are conditionally deficient inSTAT3, STAT5a/b, and JAK1 (MMTV-Cre Stat3/5 fl/fl Jak1 fl/fl ) and littermate controls without the MMTV-Cre transgene; bars, 1 mm. Dotted lines mark the invasive fronts of the terminal ends of ducts. (D) Immunoblot analysis of STAT1 expression and activation in response to the knockout of JAK1 in the mammary epithelium of STAT3/5a/5b triple-knockout females; 4 biological repeats of quadruple-knockout females compared to 2 wild-type controls and 2 STAT3/5a/5b triple-knockout mice. Beta-actin (ACTB) served as a loading control. The densitometry results of immunoblots from the 4 biological and 2 technical repeats are shown in .

Article Snippet: Rabbit polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

Techniques: Western Blot, Expressing, Activation Assay, Inhibition, Control, Quadruple Knockout, Triple Knockout, Staining, Knock-Out

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 polyclonal, pY-STAT1 (Y701) , Origene , Cat#TA309955.

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