antagonist ly341495  (Tocris)

Journal: iScience

Article Title: The maternal vGluT2 and embryonic mGluR3 signaling relay system controls offspring wing dimorphism in pea aphid

doi: 10.1016/j.isci.2025.112591

Figure Lengend Snippet: Embryonic ApmGluR3 relayed the transgenerational signal from ApvGluT2 about the induction of winged offspring (A and B) (A) Declined proportion of winged offspring by injection of an mGluR antagonist LY341495 ( n = 47), in contrast to (B) increased proportion by agonist ADPC ( n = 39). Mann-Whitney non-parametric test: ∗ p < 0.05 and ∗∗∗ p < 0.001. (C) RNAi experimental design. (D and E) (D) Knockdown of ApmGluR3 in the embryo ( n = 3) and (E) reduced proportion of winged offspring ( n = 47). (F) RNAi efficiency of ApvGluT2 in the head ( n = 4). Student’s t test was used to compare means of transcript level of ApmGluR3 . The Mann-Whitney non-parametric test was used to compare means of the proportions of winged offspring. ∗ p < 0.05 and ∗∗∗ p < 0.001. (G) Effect of ds ApvGluT2 on the proportion of winged offspring was rescued by ADPC ( n = 36). Mann-Whitney non-parametric test: ∗∗ p < 0.01. n.s. , not significant. (H) The expression of ApmGluR3 in the stage 20 embryos increased as the contact duration increased. ApmGluR3 probe conjugated with Cy5 was in red, and nuclei were stained with DAPI in blue. Wing discs shown in confocal were highlighted by white circles. (I) The relative intensity of ApmGluR3 in the wing discs of stage 20 embryos was quantified by LAS X ( n > 9). Mann-Whitney non-parametric test was used to compare the means of the proportion of winged offspring. Student’s t test was used to compare means of the relative intensity of ApmGluR3 . ∗∗ p < 0.01. n.s. , not significant. Error bars represent standard error.

Article Snippet: LY341495 , MedChemExpress , Cat#HY-70059.

Techniques: Injection, MANN-WHITNEY, Knockdown, Expressing, Staining

Journal: iScience

Article Title: The maternal vGluT2 and embryonic mGluR3 signaling relay system controls offspring wing dimorphism in pea aphid

doi: 10.1016/j.isci.2025.112591

Figure Lengend Snippet: Embryonic ApmGluR3 relayed the transgenerational signal from ApvGluT2 about the induction of winged offspring (A and B) (A) Declined proportion of winged offspring by injection of an mGluR antagonist LY341495 ( n = 47), in contrast to (B) increased proportion by agonist ADPC ( n = 39). Mann-Whitney non-parametric test: ∗ p < 0.05 and ∗∗∗ p < 0.001. (C) RNAi experimental design. (D and E) (D) Knockdown of ApmGluR3 in the embryo ( n = 3) and (E) reduced proportion of winged offspring ( n = 47). (F) RNAi efficiency of ApvGluT2 in the head ( n = 4). Student’s t test was used to compare means of transcript level of ApmGluR3 . The Mann-Whitney non-parametric test was used to compare means of the proportions of winged offspring. ∗ p < 0.05 and ∗∗∗ p < 0.001. (G) Effect of ds ApvGluT2 on the proportion of winged offspring was rescued by ADPC ( n = 36). Mann-Whitney non-parametric test: ∗∗ p < 0.01. n.s. , not significant. (H) The expression of ApmGluR3 in the stage 20 embryos increased as the contact duration increased. ApmGluR3 probe conjugated with Cy5 was in red, and nuclei were stained with DAPI in blue. Wing discs shown in confocal were highlighted by white circles. (I) The relative intensity of ApmGluR3 in the wing discs of stage 20 embryos was quantified by LAS X ( n > 9). Mann-Whitney non-parametric test was used to compare the means of the proportion of winged offspring. Student’s t test was used to compare means of the relative intensity of ApmGluR3 . ∗∗ p < 0.01. n.s. , not significant. Error bars represent standard error.

Article Snippet: Glutamate signaling agonist glutamic acid (Topscience, Shanghai) and mGluR agonist (2R, 4R)-ADPC (Enzo Life Sciences, New York) were dissolved in water at concentrations of 33.98 mM and 100 nM, respectively. vGluT2 antagonist Chicago Sky Blue 6B (CSB) (Topscience, Shanghai), mGluR antagonist LY341495 (MedChemExpress, New Jersey), and Akt inhibitor MK-2206 (Selleckchem, Houston) were dissolved in DMSO at concentrations of 10 mM, 16.98 mM, and 5 mM, respectively.

Techniques: Injection, MANN-WHITNEY, Knockdown, Expressing, Staining

Journal: Advanced Science

Article Title: GLS1‐Mediated Redundancy in Glutamate Accelerates Arterial Calcification via Activating NMDAR/Ca 2+ / β ‐Catenin Pathway

doi: 10.1002/advs.202414252

Figure Lengend Snippet: Glutamate aggravates osteogenic reprogramming of VSMCs by activating NMDAR. A) Detection of extracellular glutamate released into the supernatant of cultured VSMCs from control and Pi with or without GLS1 overexpression ( n = 3). B) Measurement of extracellular glutamate released into the supernatant of cultured VSMCs from Pi with overexpression of GLS1 in the absence or presence of DL‐TBOA (a glutamate transport inhibitor, 100 µ m ) for 7 days ( n = 5). C,D) After overexpression of GLS1, VSMCs were incubated with Pi, and then exposed to glutamate receptor inhibitors ( LY341495 : mGlu inhibitor, 10 µ m ; CNQX: AMPAR inhibitor, 10 m m ; MK801: NMDAR inhibitor, 10 µ m ), respectively. The osteogenic reprogramming of VSMCs was investigated using Alizarin Red staining ( n = 3) and Calcium quantitative analysis ( n = 3). Scale bar: 200 µm. E,F) In the presence of GLS1 overexpression, western analysis of RUNX2 was performed in VSMCs treated with Pi and 3 different glutamate receptor inhibitors ( n = 3). G,H) Dose‐response relationship for the effect of MK801 on VSMCs with overexpression of GLS1 and Pi treatment was determined by calcium content assay ( n = 3) and Alizarin Red staining ( n = 3). Scale bar: 200 µm. I,J) Western Blots analysis of VSMCs overexpressing GLS1 and treated with Pi, both in the absence and presence of MK801 ( n = 4). K) The expression of GluN1 protein levels from VSMCs in the indicated group was measured by western blot assay ( n = 5). L) Representative Von Kossa staining (upper) and immunohistochemical staining of GluN1 (lower) in arteries with or without calcification from patients ( n = 5). Scale bar: 100 µm. Data are represented as means ± SEM. One‐way ANOVA with Bonferroni's test was used in (A), (B), (D), and (J), or Dunnett's test was used in (F) and (G).

Article Snippet: Treatments were carried out simultaneously with Pi stimulation: BPTES (Selleck); CB‐839 (Selleck, 1 μ m ); MK801 (APExBIO, 10 μ m ); CNQX (APExBIO, 10 μ m ); LY341495 (APExBIO, 10 μ m ); SCH772984 (APExBIO, 10 μ m ); U0126‐EtOH (APExBIO, 10 μ m ); BAPTA‐AM (Selleck, 5 m m ); DL‐TBOA (MCE, 100 μ m ) was selected as the working concentration.

Techniques: Cell Culture, Control, Over Expression, Incubation, Staining, Western Blot, Expressing, Immunohistochemical staining