mapk pathway phosphorylation array kit (item number: aah-mapk-1-2)  (RayBiotech inc)

Journal: Disease Markers

Article Title: Knockdown of Heparanase Suppresses Invasion of Human Trophoblasts by Activating p38 MAPK Signaling Pathway

doi: 10.1155/2018/7413027

Figure Lengend Snippet: Knockdown of HPSE inhibited trophoblast cell invasion by activating p38 MAPK pathway. (a) Total cell protein including phosphorylated protein was extracted and analyzed by MAPK Pathway Phosphorylation Array C1. 1: p-mTOR. 2: p-p38 MAPK. 3: p-Rsk2. To confirm the results of microarrays, the three phosphorylated proteins and the corresponding total protein were determined in three independent experiments by Western blotting. (b) Western blotting images of p38 MAPK. (c) The gray value ratio of p-p38/t-p38 was elevated in HTR8/SVneo cells transfected with shRNA-HPSE. β -Actin was used as internal control. (d) Representative images of cell invasion after HPSE knockdown in shRNA-HPSE-HTR8 cells, pretreated with pretreated with 1 μ M BMS582949, 20 μ M SB203580, or 0.5 μ M BIRB796 for 2 h. 1: 0.1% DMSO as control. 2: 1 μ M BMS582949. 3: 20 μ M SB203580. 4: 0.5 μ M BIRB796. Magnification: 400x. (e) Quantitative analysis for the number of invasive cells. Data in graph c and graph e were represented as the mean ± SD. The differences among them were compared by one-way ANOVA and Holm-Sidak's post hoc test. ∗∗ P < 0.01.

Article Snippet: The total protein concentration was determined using a BCA assay kit (Thermo Fisher Scientific, USA), and 17 phosphorylated MAPK pathway proteins were detected semiquantitatively using a Human and Mouse MAPK Pathway Phosphorylation Array C1 Kit (Raybiotech, USA) according to the manufacturer's protocol.

Techniques: Western Blot, Transfection, shRNA

Journal: Free radical biology & medicine

Article Title: N-Alkyl triphenylvinylpyridinium conjugated dihydroartemisinin perturbs mitochondrial functions resulting in enhanced cancer versus normal cell toxicity

doi: 10.1016/j.freeradbiomed.2021.01.050

Figure Lengend Snippet: TPVP-DHA treatment of MIA PaCa-2 cells significantly decreases phosphorylation status of MAPK signaling pathways. MIA PaCa-2 cells were treated with 5 μM of TPVP-DHA for 72 h. Two hundred micrograms of total protein extracts were incubated with RayBiotec MAPK Phosphorylation Arrays following the manufacturer supplied protocol. Chemiluminescence was used to image the membrane and results were quantified using NIH ImageJ software. Images of the original blot and heat maps are shown on left and quantitative results are shown on right. Percent change in MAPK-phosphorylation status was calculated relative to MAPK-phosphorylation in cells treated with 6-TPVP alone.

Article Snippet: Cell growth and cell cycle progression were measured using the Gen5™ software as described by the manufacturer. . RayBiotec MAPK Pathway arrays: Asynchronous cultures were treated with 5 μM DHA, 6-TPVP and TPVP-DHA for 72 h. Total protein extracts were prepared following the manufacturer supplied protocol and two hundred microgram of total protein extracts were incubated with RayBiotec MAPK Pathway Phosphorylation Arrays.

Techniques: Incubation, Software