anti mypt1  (Cell Signaling Technology Inc)

Journal: Cellular Oncology

Article Title: NUAK1 silencing enhances radiotherapy-induced ferroptosis in locally advanced rectal cancer by impairing Nrf2-driven transcription of GPX4

doi: 10.1007/s13402-026-01187-5

Figure Lengend Snippet: NUAK1 knockdown combined with radiotherapy downregulates GPX4 expression by impairing Nrf2 nuclear translocation and transcriptional activity. ( A ) Dual-luciferase reporter assay measuring the activity of the GPX4 promoter with or without NUAK1 knockdown and radiation ( n = 3). ( B ) Western blot analysis of Nrf2 protein levels in nuclear and cytoplasmic fractions after NUAK1 knockdown. ( C ) Dual-luciferase reporter assay measuring the activity of ARE with or without NUAK1 knockdown and radiation ( n = 3). ( D ) Western blot analysis of protein levels of NUAK1, its downstream target (p-MYPT1), and Nrf2 downstream targets (GPX4, NQO1) under the indicated conditions. ( E ) Representative images of clonogenic survival assays under the indicated conditions after radiation. ( F ) Quantitative analysis of colony formation under the indicated conditions after radiation ( n = 4). ( G ) Representative flow cytometry histograms showing lipid peroxidation levels under the indicated conditions. ( H ) Quantitative analysis of lipid peroxidation levels under the indicated conditions ( n = 3). Data are presented as mean ± SD. * P < 0.05, **** P < 0.0001

Article Snippet: Co-precipitated proteins and input lysates were analyzed by Western blotting as described above, using antibodies against NUAK1, Nrf2, and MYPT1 (22117-1-AP, Proteintech).

Techniques: Knockdown, Expressing, Translocation Assay, Activity Assay, Luciferase, Reporter Assay, Western Blot, Flow Cytometry

Journal: Cellular Oncology

Article Title: NUAK1 silencing enhances radiotherapy-induced ferroptosis in locally advanced rectal cancer by impairing Nrf2-driven transcription of GPX4

doi: 10.1007/s13402-026-01187-5

Figure Lengend Snippet: Schematic model of the NUAK1–Nrf2–GPX4 axis in suppressing radiotherapy-induced ferroptosis in colorectal cancer. Ionizing radiation activates NUAK1, which promotes the phosphorylation of its substrate MYPT1. This signaling cascade leads to the nuclear translocation and transcriptional activation of the transcription factor Nrf2. Within the nucleus, Nrf2 binds to ARE in the promoter of the antioxidant gene, driving its transcription and increasing GPX4 protein expression. Elevated GPX4 levels inhibit lipid peroxidation, thereby suppressing ferroptosis and protecting cancer cells from radiotherapy. This diagram summarizes the key findings of this study

Article Snippet: Co-precipitated proteins and input lysates were analyzed by Western blotting as described above, using antibodies against NUAK1, Nrf2, and MYPT1 (22117-1-AP, Proteintech).

Techniques: Phospho-proteomics, Translocation Assay, Activation Assay, Expressing

Journal: Cellular Oncology

Article Title: NUAK1 silencing enhances radiotherapy-induced ferroptosis in locally advanced rectal cancer by impairing Nrf2-driven transcription of GPX4

doi: 10.1007/s13402-026-01187-5

Figure Lengend Snippet: NUAK1 knockdown combined with radiotherapy downregulates GPX4 expression by impairing Nrf2 nuclear translocation and transcriptional activity. ( A ) Dual-luciferase reporter assay measuring the activity of the GPX4 promoter with or without NUAK1 knockdown and radiation ( n = 3). ( B ) Western blot analysis of Nrf2 protein levels in nuclear and cytoplasmic fractions after NUAK1 knockdown. ( C ) Dual-luciferase reporter assay measuring the activity of ARE with or without NUAK1 knockdown and radiation ( n = 3). ( D ) Western blot analysis of protein levels of NUAK1, its downstream target (p-MYPT1), and Nrf2 downstream targets (GPX4, NQO1) under the indicated conditions. ( E ) Representative images of clonogenic survival assays under the indicated conditions after radiation. ( F ) Quantitative analysis of colony formation under the indicated conditions after radiation ( n = 4). ( G ) Representative flow cytometry histograms showing lipid peroxidation levels under the indicated conditions. ( H ) Quantitative analysis of lipid peroxidation levels under the indicated conditions ( n = 3). Data are presented as mean ± SD. * P < 0.05, **** P < 0.0001

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against NUAK1 (22723-1-AP, Proteintech), GPX4 (ab125066, Abcam), β-actin (bs-0061R, Bioss), Nrf2 (16396-1-AP, Proteintech), Lamin B1 (12987-1-AP, Proteintech), MYPT1 (22117-1-AP, Proteintech), Phospho-MYPT1 (Ser445) (CABT-BL6360, Creative Diagnostics), and NQO1 (11451-1-AP, Proteintech).

Techniques: Knockdown, Expressing, Translocation Assay, Activity Assay, Luciferase, Reporter Assay, Western Blot, Flow Cytometry

Journal: Cellular Oncology

Article Title: NUAK1 silencing enhances radiotherapy-induced ferroptosis in locally advanced rectal cancer by impairing Nrf2-driven transcription of GPX4

doi: 10.1007/s13402-026-01187-5

Figure Lengend Snippet: Schematic model of the NUAK1–Nrf2–GPX4 axis in suppressing radiotherapy-induced ferroptosis in colorectal cancer. Ionizing radiation activates NUAK1, which promotes the phosphorylation of its substrate MYPT1. This signaling cascade leads to the nuclear translocation and transcriptional activation of the transcription factor Nrf2. Within the nucleus, Nrf2 binds to ARE in the promoter of the antioxidant gene, driving its transcription and increasing GPX4 protein expression. Elevated GPX4 levels inhibit lipid peroxidation, thereby suppressing ferroptosis and protecting cancer cells from radiotherapy. This diagram summarizes the key findings of this study

Article Snippet: The membranes were then incubated overnight at 4 °C with primary antibodies against NUAK1 (22723-1-AP, Proteintech), GPX4 (ab125066, Abcam), β-actin (bs-0061R, Bioss), Nrf2 (16396-1-AP, Proteintech), Lamin B1 (12987-1-AP, Proteintech), MYPT1 (22117-1-AP, Proteintech), Phospho-MYPT1 (Ser445) (CABT-BL6360, Creative Diagnostics), and NQO1 (11451-1-AP, Proteintech).

Techniques: Phospho-proteomics, Translocation Assay, Activation Assay, Expressing

Journal: iScience

Article Title: The serotonin receptor 2b (5-HT 2B ) modulates heart remodeling following myocardial infarction via regulation of Hippo pathway

doi: 10.1016/j.isci.2026.114825

Figure Lengend Snippet: Regulation of the Hippo pathway by 5-HT 2B in cardiomyocytes (A) Western blot confirmation that Ad-FLAG-5-HT 2B infection in NRCMs resulted in the expression of FLAG- 5-HT 2B that was absent in Ad-LacZ-infected controls (representative image of n = 3 independent experiments). (B) YAP transcriptional activity was enhanced in NRCMs following 5-HT 2B overexpression ( n = 5 independent experiments with five technical replications in each experiment). (C) Representative images of GFP-YAP nuclear translocation in NRCMs overexpressing 5-HT 2B or control LacZ (scale bars, 100 μm). (D) Quantification of NRCMs with nuclear GFP-YAP showed that cells overexpressing 5-HT 2B exhibited a higher proportion of nuclear YAP ( n = 5 independent experiments with three technical replicates in each prep). (E and F) (E) Western blots and (F) quantification of band density showing a reduction in YAP phosphorylation in NRCMs overexpressing 5-HT 2B ( n = 3 independent experiments). (G and H) (G) Western blots and (H) quantification of band density showing a significant reduction in LATS1 phosphorylation in NRCMs overexpressing 5-HT 2B ( n = 3 independent experiments). (I and J) (I) Western blots and (J) quantification of band density showing no significant change in MST1 phosphorylation in NRCMs overexpressing 5-HT 2B ( n = 3 independent experiments). (K–M) (K) Western blots and (L) quantification of band density showing a reduction in expression of the LATS1 adaptor protein MOB1 in NRCMs overexpressing 5-HT 2B ( n = 3 independent experiments). (K) Western blots and (M) quantification of band density showing no change in expression of the MST adaptor protein Sav1 in NRCMs overexpressing 5-HT 2B ( n = 3 independent experiments). (N and O) (N) Western blots and (O) quantification of MYPT1 phosphorylation in NRCMs overexpressing 5-HT 2B indicated a possible role for ROCK1 linking 5-HT 2B with the Hippo pathway ( n = 3 independent experiments). (P) Immunoprecipitation analysis in cardiomyocytes overexpressing Flag-5-HT 2B using anti-Flag antibody followed by western blot detection using anti-LATS1 indicated possible interaction between 5-HT 2B and LATS1. All data are presented as mean ± SEM. Statistical tests used: (B, D, F, H, J, L, M, and O) Student’s t test.

Article Snippet: MYPT1 , Cell Signalling Technologies , Cat #2634; RRID:AB_915965.

Techniques: Western Blot, Infection, Expressing, Activity Assay, Over Expression, Translocation Assay, Control, Phospho-proteomics, Immunoprecipitation