Skip to content

Discovery and Biological Characterization of Potent MEK inhibitors in melanoma

MEK inhibitor

-tubulin was used as a loading control

Posted on March 23, 2022 By scienzaunder18

-tubulin was used as a loading control. DAPI (blue) and antibodies against Flag (red) and -tubulin (green). Control denotes expression of an irrelevant Flag-tagged protein. NIHMS6343-supplement-3.pdf (7.0M) GUID:?605D16DC-EE94-4C21-910F-42915DB736A0 4: Figure S4 A ~1.5 fold overexpression of Flag-Cep76 is sufficient to rescue centriole amplification induced by loss of Cep76 or HU treatment. (A) U2OS cells were transfected with control siRNA (siNSp) or siRNA targeting Cep76 3UTR and the indicated amounts of plasmid expressing Flag-Cep76. (Top) Western blotting of endogenous and recombinant Cep76 using an anti-Cep76 antibody. -tubulin Nedisertib was used as a loading control. (Bottom) The percentages of transfected cells with 4 CP110 dots were decided. (B) U2OS cells expressing increasing amounts of Flag-Cep76 were either left untreated or treated with HU. (Top) Western blotting of endogenous and recombinant Cep76 using an anti-Cep76 antibody. -tubulin was used as a loading control. (Bottom) The percentages of transfected cells with 4 CP110 dots were decided. In (A and B), at least 75 transfected cells were scored per condition, and common of three impartial experiments is shown. Error bars represent standard errors. Densitometric analyses were performed with ImageJ. NIHMS6343-supplement-4.pdf (438K) GUID:?026D847D-40AE-4324-A0B5-EDD543C922B2 5: Figure S5 A cancer-associated Cep76 S83C mutation is unable to suppress centriole amplification. U2OS cells transfected with control siRNA (siNSp) or siRNA targeting Cep76 3UTR (siCep76) and plasmid expressing an irrelevant Flag-tagged protein (control), Flag-Cep76 wild type (Cep76 FL) or mutant (Cep76 S83C). (A) The percentages of transfected cells with 4 CP110 dots were decided. At least 75 transfected cells were scored per condition, and average of three impartial experiments is shown. Error bars represent standard errors. (B) Cells were stained with DAPI (blue) and antibodies against Flag (red) and CP110 (green). NIHMS6343-supplement-5.pdf (1.8M) GUID:?0769A7FC-F5C1-4208-A292-6AB93952DE15 6: Figure S6 Cep76 slightly but not dramatically affects CDK2 and Cyclin A localization to the centrosome. (A) Fluorescence intensity of endogenous CDK2 at the centrosome was measured in U2OS cells transfected with control siRNA (siNSp) or Cep76 siRNA. (B) Fluorescence intensity of endogenous cyclin A at the centrosome was measured in U2OS cells transfected with control siRNA (siNSp) or Cep76 siRNA. In (A and B), at least 75 transfected cells were scored per condition, and common of three impartial experiments is shown. Error bars represent standard errors. NIHMS6343-supplement-6.pdf (342K) GUID:?4D8286E2-D20B-40E9-B535-7DA04B5964B4 7: Physique S7 Cep76 antagonizes Plk1 function to suppress centriole amplification. (A) (Left) U2OS cells were transfected with control siRNA (siNSp) or Cep76 siRNA and either left untreated or treated with BI 2536. The percentages of transfected cells with 4 centrin dots were determined. (Right) Cells were stained with DAPI (blue) and antibodies against Cep76 (red) and centrin (green). (B) U2OS cells transfected with plasmids expressing Flag and Myc, Flag-Cep76 and Myc, or Flag-Cep76 and constitutively active Plk1 (Myc-Plk1-T210D), were either left untreated or treated with HU. The percentages of transfected cells with 4 CP110 dots were decided. (ACB) At least 75 transfected cells were scored per condition, and average of three impartial experiments is shown. Error bars represent standard errors. NIHMS6343-supplement-7.pdf (723K) GUID:?C71EBC77-78C8-41AB-BECF-176B4D4D79A2 8: Figure S8 Flag-Cep76 is usually acetylated binding experiments using purified proteins revealed that while Cep76 associates weakly with cyclin E/CDK2, it binds strongly to cyclin A/CDK2 (Figure 1E), in close Nedisertib agreement with our immunoprecipitation results (Figure 1B). Taken together, these findings suggest that Cep76 preferentially interacts with cyclin A/CDK2. Open in a separate Nedisertib window Physique 1 Cep76 interacts with cyclin A/CDK2. (A) HA (control) or CDK2-HA was expressed in HEK293 cells and immunoprecipitated from lysate with an Rabbit polyclonal to LDLRAD3 anti-HA antibody. The resulting immunoprecipitates were Western blotted with anti-HA or anti-Cep76 antibodies. IN, input. (B) Flag (control) or Nedisertib Flag-Cep76 were expressed in HEK293 cells and immunoprecipitated with anti-Flag beads. Immunoprecipitates were Western blotted with the indicated antibodies. IN, input. (C) Flag or Flag-Cep76 and myc or myc-cyclin A were co-expressed in U2OS cells and Flag proteins were immunoprecipitated. Flag-Cep76 and myc-cyclin A were detected after western blotting the resulting immunoprecipitates. IN, input. (D) Western blotting of endogenous Cep76, CDK2 and Cyclin A after immunoprecipitation of U2OS cell extracts.

Oxidative Phosphorylation

Post navigation

Previous Post: The aim of the present study was to evaluate the thyroid function, throughout a specific test, in patient with RM and TAI focusing on the hypothesis that TAI should be an indirect sign of a slight thyroid dysfunction
Next Post: In addition to the DH-PH domain that is responsible for its GEF activity, Arhgef7 also contains a CH (calponin homology) and an SH3 (Src homology 3) domain at the N-terminus and proline-rich, GIT1-binding and coiled coil (CC) domains at the C-terminus (Fig

More Related Articles

Hyperkalemia is another common side effect Oxidative Phosphorylation
We display here that fumarate-mediated succination of ACO2 impairs its enzymatic activity inside a dose-dependent manner and that Fh1KO cells exhibit reduced aconitase activity Oxidative Phosphorylation

Archives

  • March 2023
  • February 2023
  • January 2023
  • December 2022
  • November 2022
  • October 2022
  • September 2022
  • August 2022
  • July 2022
  • June 2022
  • May 2022
  • April 2022
  • March 2022
  • February 2022
  • January 2022
  • December 2021
  • November 2021
  • October 2021
  • September 2021

Categories

  • Acetylcholine ??7 Nicotinic Receptors
  • Acetylcholine Nicotinic Receptors
  • Acyltransferases
  • ALK Receptors
  • Alpha1 Adrenergic Receptors
  • Angiotensin Receptors, Non-Selective
  • cMET
  • COX
  • CYP
  • Cytochrome P450
  • Decarboxylases
  • FFA1 Receptors
  • GABAA and GABAC Receptors
  • GlyR
  • H1 Receptors
  • HDACs
  • Hexokinase
  • IGF Receptors
  • K+ Ionophore
  • L-Type Calcium Channels
  • LXR-like Receptors
  • Metastin Receptor
  • Miscellaneous Glutamate
  • Neurokinin Receptors
  • Nicotinic Acid Receptors
  • Nitric Oxide, Other
  • Nucleoside Transporters
  • Opioid, ??-
  • Oxidative Phosphorylation
  • Oxytocin Receptors
  • PDK1
  • PI 3-Kinase
  • Potassium (KV) Channels
  • Potassium Channels, Non-selective
  • Prostanoid Receptors
  • Protein Kinase B
  • Protein Ser/Thr Phosphatases
  • PTP
  • Retinoid X Receptors
  • Serotonin (5-ht1E) Receptors
  • Sigma1 Receptors
  • Sirtuin
  • Syk Kinase
  • T-Type Calcium Channels
  • Transient Receptor Potential Channels
  • TRPP
  • Uncategorized
  • Urotensin-II Receptor
  • Vesicular Monoamine Transporters
  • VIP Receptors
  • XIAP

Recent Posts

  • Response to avelumab was comparable to non-binding durvalumab control conditions (Number 4F)
  • (E) Sorted we4? population
  • Journal of nuclear medicine
  • Notably, anti\SSA/Ro52 was the only auto\antibody found in almost 25% 13 of IIM individuals, both only (2/13) and in combination with anti\Jo1 (7/13), anti\PL\12 (1/13), anti\SAE (1/13), anti\SAE + anti\Jo1 (1/13), and anti\Jo1 + anti\Pm/Scl (1/13)
  • Su, H

Recent Comments

  1. A WordPress Commenter on Hello world!

Copyright © 2023 Discovery and Biological Characterization of Potent MEK inhibitors in melanoma.

Powered by PressBook Blog WordPress theme