rabbit anti phosphorylated mek pmek  (Cell Signaling Technology Inc)

Journal: Ecotoxicology and environmental safety

Article Title: Bisphenol A exposure enhances proliferation and tumorigenesis of papillary thyroid carcinoma through ROS generation and activation of NOX4 signaling pathways.

doi: 10.1016/j.ecoenv.2025.117946

Figure Lengend Snippet: Fig. 6. BPA enhances proliferation in thyroid follicular cells and BCPAP cell Xenografts in BALB/c female mice. (A) BALB/c female mice were administered either peanut oil (control) or 20 mg/kg BPA via oral gavage every other day for 4 weeks (n = 5 per group). Histological examination revealed a significant augmentation in the atypia of follicular epithelial cells in the BPA-treated group relative to the control, characterized by the presence of nuclear groove. Scale bars represent 50 μm; (B) Gross images of the extracted subcutaneous tumors are shown; (C) Tumor growth curves derived from the subcutaneous xenograft models illustrate the pro gression of tumor size over the treatment period; (D) Tumors weight of subcutaneous models; (E) The immunostain of TTF-1shows the transplanted tumor was derived from thyroid cells. Scale bars, 50 μm; (F) A partial loss of TG in BPA group. Scale bars, 100 μm; (G) Immunohistochemical staining of the tumors with or without BPA exposure is shown. Ki-67, p-AKT and p-MEK: scale bars, 50 μm; NOX4: scale bars, 100 μm.

Article Snippet: The membrane was blocked with 5 % milk powder at room temperature for 2 h, then incubated overnight at 4◦C with primary antibodies targeting, NOX4 (1:4000; Proteintech, China), phosphorylated MEK (p-MEK) (1:4000; Proteintech, China), MEK1/2 (1:20000; Proteintech, China), phosphorylated AKT (p-AKT) (1:2000; Proteintech, China), AKT (1:1000; Proteintech, China), Tubulin (1:100000; Proteintech, China), and GAPDH (1:200000; Proteintech, China).

Techniques: Control, Derivative Assay, Immunohistochemical staining, Staining

Journal: Cancer Innovation

Article Title: MAPK4 facilitates angiogenesis by inhibiting the ERK pathway in non‐small cell lung cancer

doi: 10.1002/cai2.117

Figure Lengend Snippet: Mitogen‐activated protein kinase 4 (MAPK4) silencing inhibits the proliferation of endothelial cells (ECs) by regulating the Raf/MEK/ERK1/2 signaling pathway. Human umbilical vein ECs (HUVECs) were transiently transfected with MAPK4 small interfering RNA (siRNA) (50 nM) in 24‐well plates via Lipofectamine 3000 reagent in vitro. (a) Volcano plot showing genes with differential expression in MAPK4‐silenced HUVECs (MAPK4 HUVECs) compared with NC HUVECs, as determined by RNA sequencing (RNA‐seq). n = 3 per group. (b, c) Gene set enrichment analysis plots (left) and heat maps (right) of the RNA‐seq data for NC HUVECs and MAPK4 HUVECs. (d) Western blot analysis was used to evaluate the levels of protein kinase B (AKT), phosphorylated AKT (p‐AKT), c‐Jun n‐terminal kinase (JNK), phosphorylated JNK (p‐JNK), nuclear factor κB (NF‐κB), phosphorylated NF‐κB (p‐NF‐κB), (e) ERK1/2, phosphorylated ERK1/2 (p‐ERK1/2), (f) rat sarcoma (Ras), Raf, p‐Raf, MEK, and phosphorylated MEK (p‐MEK) in HUVECs. (g) 24 h after transfection, transfected cells were treated with the p‐ERK1/2 inhibitor and cultured for another 24 h. (h) Immunofluorescence was used to evaluate and quantitatively analyze the p‐ERK1/2 level in HUVECs. Representative data from three independent experiments are shown. ** p < 0.01. ERK1/2, extracellular regulated protein kinases 1/2; MEK, mitogen‐activated extracellular signal‐regulated kinase; Raf, rapidly accelerated fibrosarcoma.

Article Snippet: Membranes were incubated with 5% skim milk in phosphate‐buffered saline (PBS) for 1 h. Immunoblotting was performed using monoclonal antibodies against cyclin‐dependent kinase inhibitor 1A (p21) (Proteintech, 10355‐1‐AP), Cyclin A (Abcam, ab185619), Cyclin B1 (Abcam, ab181593), MAPK4 (Proteintech, 26102‐1‐AP), protein kinase B (AKT) (Abcam, ab8805), phosphorylated AKT (p‐AKT) (Abcam, ab38449), ERK1/2 (Abcam, ab184699), p‐ERK1/2 (Abcam, ab201015), nuclear factor κB (NF‐κB) (Abcam, ab32536), phosphorylated NF‐κB (p‐NF‐κB) (Abcam, 97726), c‐Jun n‐terminal kinase (JNK) (Abcam, ab179461), phosphorylated JNK (p‐JNK) (Abcam, ab124956), rat sarcoma (Ras) (Abcam, ab52939), rapidly accelerated fibrosarcoma (Raf) (Abcam, ab137435), p‐Raf (Abcam, ab173539), mitogen‐activated extracellular signal‐regulated kinase (MEK) (Abcam, ab178876), phosphorylated MEK (p‐MEK) (Abcam, ab194754), and GAPDH (Abcam, 181602).

Techniques: Transfection, Small Interfering RNA, In Vitro, Expressing, RNA Sequencing Assay, Western Blot, Cell Culture, Immunofluorescence