anti phospho mapk1 erk2 p42 mapk3 erk1 p44  (Cell Signaling Technology Inc)

Journal: Cell Death & Disease

Article Title: Genes driving three-dimensional growth of immortalized cells and cancer

doi: 10.1038/s41419-025-07719-5

Figure Lengend Snippet: a The distribution of the 85 3D exclusive dropout genes in each of the top 20 molecular pathways from Fig. shown as a Matrix plot, where each purple dot represents involvement of each gene in the molecular pathways. The 24 genes shown in yellow are involved in ≥9 of the 20 molecular pathways. The red font genes are the ones which were not involved in the top 20 molecular pathways, and hence excluded from this current study. b Forest plot of the 24 genes shows the hazard ratio (HR), as well as the p value, illustrating the overall survival of BLCA patients. The blue dotted line in the middle of the plot shows an HR of 1. The 11 genes with an HR > 1 and a p value < 0.05 have the p value highlighted in red with red asterisks and written in red fonts. Genes with HR < 1 but p value < 0.05 are marked with black asterisks and written with black fonts. c Bar graph showing normalized MAPK1 gene expression across the cell transformation stages among the patient pool in GSE3167 dataset. d The expression of MAPK1 and DCLK1 in the TCGA BLCA cohort were the only two genes to show an increase among the patients who had higher stages of BLCA. e The 11 genes were assessed in the pan-cancer MetMap 500 dataset which assess the metastatic potential of cell lines, and only MAPK1 and ROCK1 showed a positive correlation with metastatic potential. f In the MetMap 500 dataset concerning exclusively BLCA cell lines, MAPK1 shows a positive correlation with a negative slope, consistent with the pan cancer analysis. The correlation is represented by a blue dotted line, which is the best fitted regression line. g In the MetMap 500 dataset concerning exclusively BLCA lines, ROCK1 does not show a positive correlation between gene effect and metastatic potential, which means that depleting ROCK1 gene in the BLCA cell lines increases the metastatic potential of the cell lines. h Radar plot shows the common metastatic sites for these BLCA cell lines, where they are likely to metastasize. i The expression level of MAPK1 in cell lines is plotted against the proliferation signature score for each cell line, with green regression line showing a positive correlation between increased MAPK1 expression and increased proliferation.

Article Snippet: These lysates were utilized for western blot of MAPK1 antibody (p-MAPK1/3 (Thr202/Tyr204; Cell Signaling Technologies #9101), following the conventional Bio-Rad SDS-PAGE protocol, in triplicates [ ].

Techniques: Gene Expression, Transformation Assay, Expressing

Journal: Cell Death & Disease

Article Title: Genes driving three-dimensional growth of immortalized cells and cancer

doi: 10.1038/s41419-025-07719-5

Figure Lengend Snippet: a The bar plot represents the expression pattern of MAPK1 across cancer types where different color gradient depict different levels of MAPK1 expression. The cancer types were abbreviated as per TCGA guidelines ( https://gdc.cancer.gov/resources-tcga-users/tcga-code-tables/tcga-study-abbreviations ). b Forest plots depict the hazard ratio for pan-cancer patients, based on their MAPK1 protein expression. The brown dotted line in the middle of the plot indicates an HR of 1. The cancer types with significant p values are colored red. The size of the square boxes corresponds to the HR value. The red boxes depict having p < 0.05, while the blue boxes are non-significant. c Overall representation of a tissue microarray slide stained with MAPK1. The normal bladder tissues containing normal urothelial layer (as defined by the commercial provider) are circled in red, whereas the cancer adjacent tissues with normal urothelium (pathologically) are circled in green. The other cores (non-circled) contain muscle layers from normal tissues and are excluded from the quantitative analysis. d Enlarged representative images of a normal urothelium core sample (upper panel) and a cancer urothelium core sample (lower panel), highlighting that both cytoplasmic and nuclear staining of MAPK1 protein was observed. Scale bar = 90 µm. e Dot plots showing the H score of each cancer core and normal core in the TMA slide. The H score provides a cumulative factor undertaking both intensity and positive staining of MAPK1 in nucleus and cytoplasm individually. f Dot plot of all BLCA cell lines, curated in DepMap analysis and correlated with MAPK1 gene expression. Note the loss of MAPK1 has little negative impact on growth of some of the BLCA cell lines (blue dots) like UMUC3 and UMUC6, while loss of MAPK1 has a dramatic negative impact on growth (red dots <−0.2) in most BLCA cell lines. g mRNA and protein levels of MAPK1 gene in UMUC3 and UMUC6 cell lines in their 2D and 3D growth condition. h Graphical representation of knocking down MAPK1 in the cancer cell lines. The adjacent western blot reveals effective knockdown of the MAPK1 in the respective cell lines. i Growth curves of UMUC3 and UMUC6 in both 2D and 3D growth conditions, where the Y axis represents CyQuant fluorescence reading. j Microscopic images of spheroids obtained by mixing UMUC3_empty vector cells labeled with DiL dye (green) with UMUC3 shMAPK1 cells labeled with DiD dye (red) at a ratio of 1:1 at day 0 (top panels). The corresponding study in UMUC6 cells is shown in the bottom panels. By day 14, both UMUC6_empty vector and UMUC3_empty vector cells dominate the population. The images shown are merged images of fluorescent field and bright field. The scale bar is 3400 μm. k Growth curves of UMUC3 empty vector and UMUC3 and UMUC6 shMAPK1 xenografts in FOXN−/− mice ( n = 5) show that both UMUC3 and UMUC6 shMAPK1 tumors grow significantly slower than empty vector control tumors. The inset picture shows representative tumors.

Article Snippet: These lysates were utilized for western blot of MAPK1 antibody (p-MAPK1/3 (Thr202/Tyr204; Cell Signaling Technologies #9101), following the conventional Bio-Rad SDS-PAGE protocol, in triplicates [ ].

Techniques: Expressing, Microarray, Staining, Gene Expression, Western Blot, Knockdown, CyQUANT Assay, Fluorescence, Plasmid Preparation, Labeling, Control

Journal: Cell Death & Disease

Article Title: Genes driving three-dimensional growth of immortalized cells and cancer

doi: 10.1038/s41419-025-07719-5

Figure Lengend Snippet: a Immunoblot showing treatment of UMUC3 and UMUC6 cells with the MAPK1 specific drug, ulixertinib, leads to decreased total MAPK1 protein, as well as decreased activated/phosphorylated MAPK1 protein (p-MAPK1), but no decrease on its homolog MAPK3. b Growth curves showing UMUC3 or UMUC6 cells treated with ulixertinib experience significant reduction in growth, relative DMSO treatment control, even at the lowest dose of drug. c HBLAK cell line treated with ulixertinib experiences significant reduction in 3D growth, compared to the DMSO treatment control, even at the lowest dose of drug, but at day 14. The scale bar is 3400 µm. d Line graph illustrating the reduction in growth of HBLAK cells after ulixertinib treatment at different timepoints. e HBLAK spheroid area, calculated from the spheroids shown in the previous figure ( c ) as well as the other biological replicates (not shown here), is plotted after ulixertinib treatment at different time points. f Schematic showing the xenograft generation and ulixertinib treatment strategy in mice ( n = 5). g Growth curves of UMUC3 and UMUC6 xenografts with ulixertinib treatment/shMAPK1 resulted in dramatic reductions, and in some case complete disappearance of UMUC3 and UMUC6 tumors.

Article Snippet: These lysates were utilized for western blot of MAPK1 antibody (p-MAPK1/3 (Thr202/Tyr204; Cell Signaling Technologies #9101), following the conventional Bio-Rad SDS-PAGE protocol, in triplicates [ ].

Techniques: Western Blot, Control