akt inhibitor api-2  (Millipore)

Journal: Zoological Research

Article Title: Red light promotes dermis-epidermis remodeling via TGFβ and AKT-mediated collagen dynamics in naturally aging mice

doi: 10.24272/j.issn.2095-8137.2024.405

Figure Lengend Snippet: Red light inhibits type I collagen degradation via AKT/NRF2/HO-1 signaling pathway activation A, B: qPCR analysis of NRF2 (A) and HO-1 (B) mRNA expression in mouse skin tissue following red light treatment. n =4. C: Representative western blot assays of p-AKT, HO-1, NRF2, and type I collagen expression in aged mice treated with red light, API-2, or both. D: Western blot analysis of nuclear-cytoplasmic separation in skin tissue. Cytosolic and nuclear extracts were immunoblotted for NRF2, while GAPDH and histone H3 proteins were probed to confirm thorough separation of the cytosolic and nuclear fractions. E: Representative western blot analysis of type I collagen expression in skin tissue after red light treatment with or without HO-1i. F: Representative immunofluorescence images of type I collagen in skin sections from aged mice after 28 days of red light treatment with or without HO-1i. Scale bar: 50 μm. G: Heatmaps showing relative expression levels of DEGs involved in various MMPs. n =3. H: qPCR analysis of MMP3, MMP9, MMP12, and MMP13 mRNA expression in mouse skin tissue following red light treatment, with and without HO-1i. *** : P <0.001; **** : P <0.0001 compared to control group, and #### : P <0.0001 compared to indicated groups, one-way ANOVA for multiple groups. Data are mean±SEM.

Article Snippet: To inhibit AKT, mice received intraperitoneal (i.p.) injections of API-2 (HY-15457, MCE, USA) at a dose of 2 mg/kg every 48 h. To inhibit TGFβ, mice received i.p. injections of SB431542 (HY-10431, MCE, USA), a TGFβR1 inhibitor, at a dose of 10 mg/kg every 48 h. To inhibit cAMP, mice received i.p. injections of SQ22536 (HY-100396, MCE, USA), an adenylate cyclase (AC) inhibitor, at a dose of 10 mg/kg every 48 h. To inhibit HO-1, mice received i.p. injections of HO-1i (HY-111798A, MCE, USA) at a dose of 1.357 mg/kg every 48 h. To inhibit intracellular free Ca 2+ , mice received i.p. injections of diltiazem (HY-B0632, MCE, USA) at a dose of 20 mg/kg every 48 h. To inhibit reactive oxygen species (ROS), mice received daily topical application of 100 mg/mL ascorbic acid (HY-B0166, MCE, USA), applied to the skin 30 min after red light exposure.

Techniques: Activation Assay, Expressing, Western Blot, Immunofluorescence, Control

Journal: International Journal of Molecular Medicine

Article Title: Curcumin attenuates myocardial ischemia-reperfusion-induced autophagy-dependent ferroptosis via Sirt1/AKT/FoxO3a signaling

doi: 10.3892/ijmm.2025.5492

Figure Lengend Snippet: Cur decreases A/R injury in H9c2 cells via Sirt1. Expression of (A) apoptosis-associated proteins in A/R-injured H9c2 cells following Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity. (B) Relative protein expression of Bcl2. (C) represents the relative protein expression of Bax. Expression of Sirt1 (D) in A/R-injured H9c2 cells following Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity. (E) Relative protein expression of Sirt1. Apoptosis rate of H9c2 cells was measured by flow cytometry (F and G) illustrates the proportion of apoptotic cells as determined by flow cytometry. (H) caspase 3, (I) LDH activity and (J) viability of A/R injured H9c2 cells after Cur pretreatment, silencing of Sirt1 expression and targeted inhibition of AKT activity. * P<0.05, ** P<0.01, *** P<0.001. Cur, curcumin; A/R, anoxia/reoxygenation; Sirt, silent information regulator 1; LDH, Lactate dehydrogenase; si, small interfering; NC, non-targeting control; API-2, triciribine; CON, control.

Article Snippet: Cur (purity ≥98%, batch no. DC0279-0005) was purchased from Dester Technology Co., Ltd. and Akt inhibitor triciribine (API-2; cat. no. GC15392) was purchased from GLPBIO Technology LLC.

Techniques: Expressing, Inhibition, Activity Assay, Flow Cytometry, Control

Journal: International Journal of Molecular Medicine

Article Title: Curcumin attenuates myocardial ischemia-reperfusion-induced autophagy-dependent ferroptosis via Sirt1/AKT/FoxO3a signaling

doi: 10.3892/ijmm.2025.5492

Figure Lengend Snippet: Cur reduces autophagy-dependent ferroptosis in H9c2 cells associated with A/R injury via Sirt1. Expression of autophagy-(A) and ferroptosis-related proteins (B) in A/R-injured H9c2 cells following Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity. (C) Relative protein expression of P62 and the ratio of LC3II/I. (D) Relative protein expression of NCOA4 and FTH1. Detection of (E) total iron ions, (F) MDA, (G) GSSG, (H) GSH, (I) GSH/GSSG and (J) SOD in A/R injured H9c2 cells following Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity. Fluorescence intensity of (K) lysosomes, (L) ROS and (M) Fe 2+ in A/R-injured H9c2 cells following Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity (magnification, ×200; scale bar, 400 μ m). ** P<0.05, *** P<0.01. Cur, curcumin; A/R, anoxia/reoxygenation; Sirt, silent information regulator 1; MDA, malondialdehyde; GSSG, glutathione disulfide; GSH, glutathione; SOD, superoxide dismutase; ROS, reactive oxygen species; NCOA4, nuclear receptor coactivator 4; FTH1, ferritin heavy chain 1; CON, control; si, small interfering; prot, protein; API, triciribine; LC3II, microtubule-associated protein 1 light chain 3 β; NC, non-targeting control.

Article Snippet: Cur (purity ≥98%, batch no. DC0279-0005) was purchased from Dester Technology Co., Ltd. and Akt inhibitor triciribine (API-2; cat. no. GC15392) was purchased from GLPBIO Technology LLC.

Techniques: Expressing, Inhibition, Activity Assay, Fluorescence, Control

Journal: International Journal of Molecular Medicine

Article Title: Curcumin attenuates myocardial ischemia-reperfusion-induced autophagy-dependent ferroptosis via Sirt1/AKT/FoxO3a signaling

doi: 10.3892/ijmm.2025.5492

Figure Lengend Snippet: Cur mediates nuclear localization of FoxO3a via Sirt1/AKT. (A) Western blot analysis of (B) phosphorylation of AKT and FoxO3a in A/R-injured H9c2 cells after Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity. (C) Western blot analysis of (D) distribution of FoxO3a in the cytoplasm and nucleus of A/R-injured H9c2 cells following Cur pretreatment, Sirt1 silencing and targeted inhibition of AKT activity. *** P<0.01. Cur, curcumin; PCNA, proliferating cell nuclear antigen; Sirt, silent information regulator 1; A/R, anoxia/reoxygenation; p-, phosphorylated; si, small interfering; NC, non-targeting control; API, Triciribine; CON, control.

Article Snippet: Cur (purity ≥98%, batch no. DC0279-0005) was purchased from Dester Technology Co., Ltd. and Akt inhibitor triciribine (API-2; cat. no. GC15392) was purchased from GLPBIO Technology LLC.

Techniques: Western Blot, Inhibition, Activity Assay, Control