targeting erk1 (Cell Signaling Technology Inc)

Cell Signaling Technology Inc targeting erk1

A, The phosphorylation of <t>ERK1/2</t> in different cancer cells. Cancer cells were treated with NSC59984 (μmol/L) for 16 hours. B. The phosphorylation of ERK1/2 in SW480 cells treated with NSC59984 (μmol/L) in a dose and time course. C, The phosphorylation of ERK1/2 in SW480 cancer cells treated with NSC59984 (μmol/L) and NAC (5 mmol/L) for 16 hours. D, The phosphorylation of ERK1/2 in SW480 cancer cells with knockdown of Ras. SW480 cells were transfected with siRNAs (siRNA #1 and #2) to knockdown Ras, followed with NSC59984 (μmol/L) treatment for 16 hours. E, The phosphorylation of ERK1/2 in SW480 cells with knockdown of Raf expression. C-Raf or B-Raf expression was knocked down by siRNA in SW480 cells, followed with NSC59984 (μmol/L) treatment for 16 hours. F, The phosphorylation of ERK1/2 in SW480 cells treated with NSC59984 (μmol/L) and the different kinase inhibitors (μmol/L) as indicated for 16 hours.

Targeting Erk1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A, The phosphorylation of ERK1/2 in different cancer cells. Cancer cells were treated with NSC59984 (μmol/L) for 16 hours. B. The phosphorylation of ERK1/2 in SW480 cells treated with NSC59984 (μmol/L) in a dose and time course. C, The phosphorylation of ERK1/2 in SW480 cancer cells treated with NSC59984 (μmol/L) and NAC (5 mmol/L) for 16 hours. D, The phosphorylation of ERK1/2 in SW480 cancer cells with knockdown of Ras. SW480 cells were transfected with siRNAs (siRNA #1 and #2) to knockdown Ras, followed with NSC59984 (μmol/L) treatment for 16 hours. E, The phosphorylation of ERK1/2 in SW480 cells with knockdown of Raf expression. C-Raf or B-Raf expression was knocked down by siRNA in SW480 cells, followed with NSC59984 (μmol/L) treatment for 16 hours. F, The phosphorylation of ERK1/2 in SW480 cells treated with NSC59984 (μmol/L) and the different kinase inhibitors (μmol/L) as indicated for 16 hours.

Journal: Molecular cancer research : MCR

Article Title: Small-molecule NSC59984 induces mutant p53 degradation through a ROS-ERK2-MDM2 axis in cancer cells

doi: 10.1158/1541-7786.MCR-21-0149

Figure Lengend Snippet: A, The phosphorylation of ERK1/2 in different cancer cells. Cancer cells were treated with NSC59984 (μmol/L) for 16 hours. B. The phosphorylation of ERK1/2 in SW480 cells treated with NSC59984 (μmol/L) in a dose and time course. C, The phosphorylation of ERK1/2 in SW480 cancer cells treated with NSC59984 (μmol/L) and NAC (5 mmol/L) for 16 hours. D, The phosphorylation of ERK1/2 in SW480 cancer cells with knockdown of Ras. SW480 cells were transfected with siRNAs (siRNA #1 and #2) to knockdown Ras, followed with NSC59984 (μmol/L) treatment for 16 hours. E, The phosphorylation of ERK1/2 in SW480 cells with knockdown of Raf expression. C-Raf or B-Raf expression was knocked down by siRNA in SW480 cells, followed with NSC59984 (μmol/L) treatment for 16 hours. F, The phosphorylation of ERK1/2 in SW480 cells treated with NSC59984 (μmol/L) and the different kinase inhibitors (μmol/L) as indicated for 16 hours.

Article Snippet: Anti-cleaved caspase 3 (catalog no. 559565) was purchased from BD Pharmingen. siRNA#1 targeting ERK2 (s11138, catalog no.4390824), siRNA#2 targeting ERK2 (s11139, catalog no.4390824), siRNA#1 targeting MDM2 (s8628, no.4390824), siRNA#2 targeting MDM2 (s8630, catalog no.4390824), siRNA#2 targeting ERK1(s11140, catalog no.4390824) and siRNA#2 targeting Ras (s7939, catalog no.4390824) were purchased from Ambion. siRNA#1 targeting ERK1 (catalog no. 6436) were purchased from Cell Signaling Technology. siRNA#1 targeting Ras (catalog no. sc-35731), siRNA targeting C-Raf (catalog no.sc-29462) and siRNAs targeting B-Raf (catalog no. sc-36368) were purchased from Santa Cruz Biotechnology.

Techniques: Phospho-proteomics, Knockdown, Transfection, Expressing

A, The expression of mutant p53 at the protein level in SW480 cells treated with NSC59984 (μmol/L) and NAC (mmol/L) for 16 hours. B, The expression of mutant p53 at the protein level in SW480 cancer cells treated with NSC59984 (μmol/L) and U0126 (10 μmol/L) or Sorafenib (32 μmol/L) for 16 hours. C, The expression of mutant p53, p21 and noxa at the protein levels in SW480 cancer cells treated with NSC59984 (μmol/L) and ERK1/2 inhibitor, SCH772984 (SCH, 1μmol/L) or VX-11e (5 μmol/L) for 16 hours. D, The ubiquitination assay by IP in SW480 cells. SW480 cells were transfected with HA-Ub for 48 hours, followed with the treatment with NSC59984 (μmol/L) and U0126 (10 μmol/L) for an additional 8 hours. E, The protein level of mutant p53 and p53 targets in SW480 cells treated with NSC59984 (μmol/L) and SP600125 (10 μmol/L) for 16 hours. F, The expression of mutant p53 at the protein level in the cancer cells with knockdown of Ras and ERK2. Ras and ERK2 were knocked down in SW480 cells with siRNA (#1 and #2 for each target), followed by treatment with NSC59984 (μmol/L) and 20 μmol/L of Z-VAD-FMK for 16 hours. G, The expression of the mutant p53 at the protein level in the ERK1-knockdown SW480 cells treated with NSC59984 (μmol/L) for 16 hours. ERK1 was knocked down by two siRNAs. H, The expression of the mutant p53 at the protein level in the Raf-knockdown cancer cells. C-Raf and B-Raf were knocked down in SW480 cells by siRNA, followed with NSC59984 treatment (μmol/L) for 16 hours.

Journal: Molecular cancer research : MCR

Article Title: Small-molecule NSC59984 induces mutant p53 degradation through a ROS-ERK2-MDM2 axis in cancer cells

doi: 10.1158/1541-7786.MCR-21-0149

Figure Lengend Snippet: A, The expression of mutant p53 at the protein level in SW480 cells treated with NSC59984 (μmol/L) and NAC (mmol/L) for 16 hours. B, The expression of mutant p53 at the protein level in SW480 cancer cells treated with NSC59984 (μmol/L) and U0126 (10 μmol/L) or Sorafenib (32 μmol/L) for 16 hours. C, The expression of mutant p53, p21 and noxa at the protein levels in SW480 cancer cells treated with NSC59984 (μmol/L) and ERK1/2 inhibitor, SCH772984 (SCH, 1μmol/L) or VX-11e (5 μmol/L) for 16 hours. D, The ubiquitination assay by IP in SW480 cells. SW480 cells were transfected with HA-Ub for 48 hours, followed with the treatment with NSC59984 (μmol/L) and U0126 (10 μmol/L) for an additional 8 hours. E, The protein level of mutant p53 and p53 targets in SW480 cells treated with NSC59984 (μmol/L) and SP600125 (10 μmol/L) for 16 hours. F, The expression of mutant p53 at the protein level in the cancer cells with knockdown of Ras and ERK2. Ras and ERK2 were knocked down in SW480 cells with siRNA (#1 and #2 for each target), followed by treatment with NSC59984 (μmol/L) and 20 μmol/L of Z-VAD-FMK for 16 hours. G, The expression of the mutant p53 at the protein level in the ERK1-knockdown SW480 cells treated with NSC59984 (μmol/L) for 16 hours. ERK1 was knocked down by two siRNAs. H, The expression of the mutant p53 at the protein level in the Raf-knockdown cancer cells. C-Raf and B-Raf were knocked down in SW480 cells by siRNA, followed with NSC59984 treatment (μmol/L) for 16 hours.

Techniques: Expressing, Mutagenesis, Ubiquitin Proteomics, Transfection, Knockdown

A, The protein level of mutant p53 and p53 targets in HT29 cells following treatment with NSC59984 (μmol/L) in combination with NAC (10 mmol/L) or BSO (10 μmol/L) for 16 hours. B, p53-responsive reporter bioluminescence in SW480 cells treated with NSC59984 (μmol/L) and NAC (10 mmol/L) for 16 hours. C, p53 responsive reporter bioluminescence in SW480 cells treated with NSC59984 (μmol/L) and BSO (10 μmol/L) for 16 hours. D, The expression of mutant p53 and p53 targets at the protein level in SW480 cells treated with NSC59984 (μmol/L) and U0126 (10 μmol/L) for 16 hours. E, The protein level of mutant p53 and p53 targets in RXF393 cancer cells carrying mutant p53 (R175H). The cells were treated with NSC59984 (μmol/L) and U0126 (10 μmol/L) or BSO (20 μmol/L) for 16 hours. F, The expression of mutant p53 and p53 targets at the protein levels in HT29 cells treated with NSC59984 (μmol/L) and ERK1/2 inhibitor SCH772984 (1μmol/L) or VX-11e (5μmol/L) for 16 hours. G, The p53-responsive reporter bioluminescence assay in SW480 cells with the overexpression of Ad-p73. p73 was overexpressed in SW480 cells by adenovirus (Ad-p73) infection. The cells were treated with NSC59984 (μmol/L) and U0126 (10μmol/L) or SP600125 (10μmol/L) for 16 hours. Data represent mean ± SD. *, P<0.05 compared with the NSC59984 treatment at each dosage. H, The Ad-p73 expressed protein levels in the cells with the adenovirus infection(G). I, ChIP-PCR assay. p73 was overexpressed in HT29 with adenovirus infection. Following treatment with NSC59984 (μmol/L) and U0126 (μmol/L) for 7 hours. ChIP assay was performed with anti-P73 and IgG as a control. The p21 or Noxa promoters were quantified by real-time PCR using the ChIP-eluted DNA. Data were normalized to the ChIP with anti-p73 in the cells treated with DMSO treatment as a control. Data represent mean ± SD, N=2. ANOVA test, P<0.05. J. P21 and Noxa at the protein level in HT29 cells by Western blot assay (I).

Journal: Molecular cancer research : MCR

Article Title: Small-molecule NSC59984 induces mutant p53 degradation through a ROS-ERK2-MDM2 axis in cancer cells

doi: 10.1158/1541-7786.MCR-21-0149

Figure Lengend Snippet: A, The protein level of mutant p53 and p53 targets in HT29 cells following treatment with NSC59984 (μmol/L) in combination with NAC (10 mmol/L) or BSO (10 μmol/L) for 16 hours. B, p53-responsive reporter bioluminescence in SW480 cells treated with NSC59984 (μmol/L) and NAC (10 mmol/L) for 16 hours. C, p53 responsive reporter bioluminescence in SW480 cells treated with NSC59984 (μmol/L) and BSO (10 μmol/L) for 16 hours. D, The expression of mutant p53 and p53 targets at the protein level in SW480 cells treated with NSC59984 (μmol/L) and U0126 (10 μmol/L) for 16 hours. E, The protein level of mutant p53 and p53 targets in RXF393 cancer cells carrying mutant p53 (R175H). The cells were treated with NSC59984 (μmol/L) and U0126 (10 μmol/L) or BSO (20 μmol/L) for 16 hours. F, The expression of mutant p53 and p53 targets at the protein levels in HT29 cells treated with NSC59984 (μmol/L) and ERK1/2 inhibitor SCH772984 (1μmol/L) or VX-11e (5μmol/L) for 16 hours. G, The p53-responsive reporter bioluminescence assay in SW480 cells with the overexpression of Ad-p73. p73 was overexpressed in SW480 cells by adenovirus (Ad-p73) infection. The cells were treated with NSC59984 (μmol/L) and U0126 (10μmol/L) or SP600125 (10μmol/L) for 16 hours. Data represent mean ± SD. *, P<0.05 compared with the NSC59984 treatment at each dosage. H, The Ad-p73 expressed protein levels in the cells with the adenovirus infection(G). I, ChIP-PCR assay. p73 was overexpressed in HT29 with adenovirus infection. Following treatment with NSC59984 (μmol/L) and U0126 (μmol/L) for 7 hours. ChIP assay was performed with anti-P73 and IgG as a control. The p21 or Noxa promoters were quantified by real-time PCR using the ChIP-eluted DNA. Data were normalized to the ChIP with anti-p73 in the cells treated with DMSO treatment as a control. Data represent mean ± SD, N=2. ANOVA test, P<0.05. J. P21 and Noxa at the protein level in HT29 cells by Western blot assay (I).

Techniques: Mutagenesis, Expressing, ATP Bioluminescent Assay, Over Expression, Infection, Control, Real-time Polymerase Chain Reaction, Western Blot

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