Journal: Molecular Therapy

Article Title: Erythropoietin Gene-enhanced Marrow Mesenchymal Stromal Cells Decrease Cisplatin-induced Kidney Injury and Improve Survival of Allogeneic Mice

doi: 10.1038/mt.2011.162

Figure Lengend Snippet: Effect of conditioned media (CM) from mesenchymal stromal cells (MSCs) and from Epo-MSCs on P-Akt expression in kidney cells. MM55.K mouse kidney epithelial cells were exposed to cisplatin, with/without MSCs CM or Epo-MSCs CM (a) at increasing concentrations or (b) at highest concentrations with/without a neutralizing anti-IGF-1 antibody (IGF1 Ab), and/or anti-VEGF antibody (VEGF Ab), or anti-Epo antibody (Epo Ab), as detailed in the Materials and Methods. Cells were collected 42 hours later and lysates used for western blot analysis of phosphorylated Akt (Ser473) (P-Akt) expression, as well as of loading controls GAPDH or total Akt expression. (b) The bar graph represents the mean ± SEM of seven independent experiments.

Article Snippet: The Epo-MSCs CM was similarly tested alone or in combination with neutralizing antibodies against IGF-1, VEGF or Epo (MAB959) (R&D Systems), or against both IGF-1and VEGF.

Techniques: Expressing, Western Blot

Journal: Lipids in Health and Disease

Article Title: Deficiency of lysophosphatidic acid receptor 3 decreases erythropoietin production in hypoxic mouse kidneys

doi: 10.1186/s12944-024-02367-8

Figure Lengend Snippet: LPAR3 regulates the HIF-2α‒EPO axis in hypoxic Hep3B cells. A-D Hep3B cells were transfected with nonspecific siRNA (NC siRNA), LPAR3 siRNA-1 or LPAR3 siRNA-2 for 48 h and then exposed to 1% O 2 for 12 h. LPAR3 mRNA (A) and EPO mRNA expression levels (B) were determined via RT‒qPCR, EPO protein levels in culture medium (C) were determined via ELISAs, and HIF-2α protein levels in Hep3B cells (D) were determined via Western blotting. E-G Hep3B cells were starved in serum-free MEM for 24 h, pretreated in the presence or absence of 30 µM Ki16425 for 20 min, and then challenged with 10 µM 2S-OMPT for 1 h, followed by hypoxia exposure (1% O 2 ) for 6 h. EPO mRNA expression levels (E) were determined via RT‒qPCR, EPO protein levels in culture medium (F) were determined via ELISAs, and HIF-2α protein levels in Hep3B cells (G) were determined via Western blotting. n = 3 in each group. The p value was determined by 2-way analysis of variance (ANOVA) followed by a post hoc test. *** p < 0.001, **** p < 0.001 relative to the normoxia control of the same treatment; ## p < 0.01, ### p < 0.001, #### p < 0.0001, between different siRNA-treated or different drug-treated groups; mean ± SD

Article Snippet: EPO levels in mouse plasma were determined by ELISAs (Cat No: KE10031; Proteintech, IL, USA).

Techniques: Transfection, Expressing, Western Blot, Control

Journal: Lipids in Health and Disease

Article Title: Deficiency of lysophosphatidic acid receptor 3 decreases erythropoietin production in hypoxic mouse kidneys

doi: 10.1186/s12944-024-02367-8

Figure Lengend Snippet: LPAR3 promotes the HIF-2α‒EPO axis via the PI3K‒AKT pathway. A Heatmap of the downregulated genes enriched in the PI3K‒Akt signaling pathway in the kidneys of hypoxic mice with LPAR3 deficiency. B The total renal AKT and p-AKT levels in the WT and Lpar3 −/− mice under normoxic or hypoxic conditions (8% O 2 ) were determined by Western blotting, n = 6. The p value was determined by 2-way analysis of variance (ANOVA) followed by a post hoc test. *** p < 0.001 compared with the isogenic normoxia control; # p < 0.05 between different genotype groups; mean ± SEM. C Hep3B cells were transfected with nonspecific siRNA (NC siRNA), LPAR3 siRNA-1 or LPAR3 siRNA-2 for 48 h and then exposed to 1% O 2 for 12 h. Total AKT and phosphorylated AKT (p-AKT) levels were determined via Western blotting. D-F Hep3B cells were starved in serum-free MEM for 24 h, pretreated with or without LY294002 (10 µM) for 20 min, and then challenged with 2S-OMPT for 1 h, followed by hypoxia exposure (1% O 2 ) for 6 h. Total AKT, p-AKT, and HIF-2α protein levels were determined by Western blotting (D) . The EPO mRNA expression levels (E) were determined via RT‒qPCR, and the EPO protein levels in the culture medium (F) were determined via ELISAs. The p value was determined by 2-way analysis of variance (ANOVA) followed by a post hoc test. ** p < 0.01, **** p < 0.0001 relative to the normoxia control of the same treatment; ### p < 0.001, #### p < 0.0001, between different drug-treated groups; mean ± SD

Article Snippet: EPO levels in mouse plasma were determined by ELISAs (Cat No: KE10031; Proteintech, IL, USA).

Techniques: Western Blot, Control, Transfection, Expressing