akt inhibitor api 2  (Tocris)

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