Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: Granulosa cells collected by follicular puncture from mice treated with PD0325901 showed absence of phosphorylation of Mapk3/1 at Thr302/Tyr204 compared to vehicle treated mice (n = 3 mice/group/time point).

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques: Phospho-proteomics

Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: Treatment of immature mice with single dose of 25μg/g of PD0325901 resulted in anovulation compared to vehicle treated mice (A) during superovulation (n = 5 mice/ group). This anovulation as evidenced by trapped GV stage oocytes with compact cumulus cells (C&D) inside the preovulatory follicles at 18h hCG (n = 5 mice/ group). Vehicle treated mice showed well-developed corpus luteum (B) evidencing follicular rupture, ovulation and luteinization. CL—corpus luteum; GV—germinal vesicle.

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques:

Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: Treatment of immature mice with single dose of 25μg/g of PD0325901 did not alter relative mRNA abundance of Mapk1 and Mapk3 (A). Expression of known hCG regulated genes like Fshr and Nr5a2 (B) as well as hCG induced genes like Scarb1 and Pappa (C) were also not altered in PD0325901 treated mice (n = 3mice/group/time point).

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: Treatment of immature mice with single dose of 25μg/g of PD0325901 inhibited the expression of hCG-induced genes involved in follicular rupture like Ptgs2 , Pgr , Adamts1 ; genes involved in cumulus expansion like Has2 , Ptx3 , Tnfaip6 ; gene involved in oocyte meiotic maturation like Areg and luteinization like Cebpb (n = 3mice/group/time point). Mapk3/1 regulated LH induced genes like Ptgs2 , Ptx3 and Tnfaip6 were not regulated by Cebpb.

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: A) Treatment of immature mice with single dose of 25μg/g of PD0325901 inhibited the expression of hCG-induced expression of Egr1 1h and 4h hCG. B) Abundance of Egr1 protein in granulosa cells of the inhibitor and vehicle treated mice. Actb was used as loading control.

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques: Expressing, Control

Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: A) Treatment with PD0325901 decreased Fo+PMA induced increases in the abundance of Ptgs2 protein relative to vehicle treated primary granulosa cells. B) Pre-treatment with Egr1-siRNA inhibited Fo+PMA induced expression of Egr1 when compared to control siRNA treatment in primary granulosa cells in vitro. This was associated with reduced abundance of Pgts2 transcript in Egr1-siRNA treated cells. Fo—forskolin; PMA—phorbol-12-myristate.

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques: Expressing, Control, In Vitro

Journal: PLoS ONE

Article Title: Effect of the Transient Pharmacological Inhibition of Mapk3/1 Pathway on Ovulation in Mice

doi: 10.1371/journal.pone.0119387

Figure Lengend Snippet: Preovulatory LH surge induced Mapk3/1 activity regulates transcription of ovulatory genes such as Ptgs2 trough transcription factor Egr1. Inhibition of this LH-induced Mapk3/1 activity by Map2k inhibitor PD0325901 results in anovulation.

Article Snippet: A potent selective Map2k (MEK) inhibitor PD0325901 (Selleckchem) was initially dissolved in DMSO (Fisher Scientific) to prepare a stock solution of 100 μg/μl concentration.

Techniques: Activity Assay, Inhibition

Journal: Disease Models & Mechanisms

Article Title: MEK-inhibitor-mediated rescue of skeletal myopathy caused by activating Hras mutation in a Costello syndrome mouse model

doi: 10.1242/dmm.049166

Figure Lengend Snippet: Treatment of primary myoblast cultures with MEK inhibitor (MEKi). Primary Hras G12V and WT myoblast cultures were treated with and without the addition of MEKi PD0325901 to a final concentration of 1 µM. (A) Merged images of immunofluorescently labeled MyHC-expressing cells (green) with DAPI-counterstained nuclei (blue) (10× magnification). (B) At 24 h (left) and 48 h (right) in DM with no MEKi, control Hras G12V cultures had significantly fewer nuclei in MyHC-expressing differentiated cells, indicating less myotube formation than in WT cultures ( n =5; P <0.01). With the addition of MEKi, the Hras G12V cultures showed a significant increase in the number of nuclei found in MyHC-expressing differentiated cells and concomitantly showed an increase in myotube formation compared to the untreated Hras G12V cultures ( n =5; P <0.01). MEKi-treated Hras G12V cultures exhibited robust elongated myotube formation by 48 h in DM. Importantly, at both 24 h and 48 h in DM, the number of nuclei in MyHC-expressing differentiated cells in the MEKi-treated Hras G12V cultures was not significantly different from that in either the WT control or WT MEKi cultures ( P =0.45 at 24 h and P =0.16 at 48 h, one-way ANOVA). The MEKi did not significantly alter the differentiation process in the WT myoblast culture. The bars represent the mean±s.e.m. ** P <0.01; NS, not significant (unpaired Student's t -test).

Article Snippet: Proliferating myoblasts from Hras G12V and WT cultures were incubated in differentiation medium (DM), with or without the addition of MEK (also known as MAP2K) inhibitor (MEKi) PD0325901 (Pfizer; 1 μM final concentration).

Techniques: Concentration Assay, Labeling, Expressing, Control

Journal: Cell Death Discovery

Article Title: A six-inhibitor culture medium for improving naïve-type pluripotency of porcine pluripotent stem cells

doi: 10.1038/s41420-019-0184-4

Figure Lengend Snippet: a Live cell images of H1 hESC and Lv-piPSC under F and NHSM conditions with bars, 200 µm and AP staining images in NHSM condition with bars, 5 mm. b AP staining images of LV-piPSC following removal of individual components from NHSM. AP staining was performed at Day 5 and Day 10 of culture under each condition with bar, 5 mm. c Live cell (bar, 500 µm) and AP staining (bar, 5 mm) images of Lv-piPSC under NHSM, NHSM without TGFB1 and NHSM without TGFB1, plus TGFB inhibitor (TGFBi) conditions. d AP staining images of Lv-piPSC under FL6i condition and FL6i in absence of respective components (bar, 500 µm)

Article Snippet: Cytokine and small molecules include 20 ng/ml hLIF (Millipore), 8 ng/ml hFGF2, 2 ng/ml TGFB1 (Prospec), 2 μM p38i (MAPK14 inhibitor, BIRB796, Selleckchem), 5 μM JNKi (MAPK8 inhibitor, SP600125, Tocris), 0.4 μM BMP inhibitor (LDN193189, Axon), 3 μM GSK3A inhibitor (CHIR99021, STEMCELL Technologies), and 1 μM ERK1/2i (MAP2K inhibitor, PD0325901, Selleckchem) .

Techniques: Staining