anti-pmek (41g9)  (Cell Signaling Technology Inc)

Journal: Signal Transduction and Targeted Therapy

Article Title: Tet methylcytosine dioxygenase 2 (TET2) deficiency elicits EGFR-TKI (tyrosine kinase inhibitors) resistance in non-small cell lung cancer

doi: 10.1038/s41392-024-01778-4

Figure Lengend Snippet: TET2 deficiency elicits the activation of TNF/NF-κB signaling. Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) analyses of upregulated differential genes in HCC827-OR cells ( a ) and HCC827 TET2-KO ( b ) cells compared with HCC827 naïve cells. c Scatter plots to show the differential gene expression and the differential hypomethylated regions in gene body derived from integrated analysis of RNA-Seq and 5hmC-Seq data in HCC827 and HCC827 TET2-KO cells. d KEGG analyses of genes within differential hypomethylated regions in gene body in HCC827 TET2-KO cells, the gene sets analyzed are adapted from c shown in blue color. Enzyme-linked immunosorbent assay (ELISA) to measure the TNFα ( e ) and IL-6 ( f ) level in indicated cell lines. g IB analyses of pRelA S536 , RelA and IκBα in the cell lines described in ( f ). h ELISA to measure TNFα and IL-6 level in PC-9OR or HCC827-OR cells with or without TET2 S1107 overexpression. i IB analyses of pRelA S536 , RelA and IκBα in the cell lines described in ( h ). j Representative IHC staining of pMEK, TET2, and RelB in specimens from NSCLC patients and PDXs, which were resistant (Patient-1R, PDX-1R) or sensitive (Patient-1S, PDX-1S) to osimertinib treatment. Scale bars, 50 μm. P values were calculated using two-tailed unpaired Student’s t tests ( e , f , h ). Data in ( e – i ) are representative of two independent experiments. Data are shown as mean ± SEM

Article Snippet: Subsequently, sections were blocked with goat serum and incubated overnight with primary antibodies against pMEK (CST, #9154, 1:200), TET2 (CST, #18950, 1:100), or RelB (HuaBio, ET1612-18, 1:100).

Techniques: Activation Assay, Gene Expression, Derivative Assay, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Over Expression, Immunohistochemistry, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Tet methylcytosine dioxygenase 2 (TET2) deficiency elicits EGFR-TKI (tyrosine kinase inhibitors) resistance in non-small cell lung cancer

doi: 10.1038/s41392-024-01778-4

Figure Lengend Snippet: Working model. This model depicts the pMEK, TET2 and NF-κB expression patterns in EGFR mut cancer cells treated by EGFR-TKI. Short-term exposed to EGFR-TKI inhibit MEK activity and downregulate TET2 whereas the NF-κB has not been evoked in EGFR mut cancer cells. Nevertheless, various TKI-resistant mechanisms were reported and can be summarized in at least three aspects: ( a ) MEK reactivation, with the most cases of amplifications, fusions or mutations among RTK-RAS-MEK pathway; ( b ) NF-κB activation, featured by sustained MEK1 inhibition and the following TET2 low expression and NF-κB activation. c Other mechanisms, like EMT or histological transformation. The picture at the right was a further illustration of ( b ). In brief, the loss of TET2 mediated by MEK1 inactivation resulted in the upregulation of NF-κB pathway thus elicits resistance to EGFR-TKIs. This figure includes a portion generated from BioRender ( https://app.biorender.com/ )

Article Snippet: Subsequently, sections were blocked with goat serum and incubated overnight with primary antibodies against pMEK (CST, #9154, 1:200), TET2 (CST, #18950, 1:100), or RelB (HuaBio, ET1612-18, 1:100).

Techniques: Expressing, Activity Assay, Activation Assay, Inhibition, Transformation Assay, Generated