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miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor <t>DAPT</t> (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.
γ Secretase Inhibitor Dapt, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor <t>DAPT</t> (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.
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miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor <t>DAPT</t> (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.
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miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor <t>DAPT</t> (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.
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miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor <t>DAPT</t> (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.
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miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor DAPT (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.

Journal: International Journal of Molecular Medicine

Article Title: miR-223-3p promotes microglial lactylation and M1 polarization via the FBXW7/Notch1/Hes1/SIRT1 axis

doi: 10.3892/ijmm.2026.5849

Figure Lengend Snippet: miR-223-3p suppresses SIRT1 transcription via the Notch1/Hes1 signaling pathway, thereby promoting microglial lactylation and M1 polarization. (A) RT-qPCR analysis of Notch1 and Hes1 mRNA levels. (B) Western blot analysis of NICD1 and Hes1 protein expression. (C) Western blot analysis of SIRT1 expression following silencing Hes1. (D) Western blot detection of global lactylation levels. (E) RT-qPCR analysis of microglial polarization markers. (F and G) RT-qPCR and western blot detection of SIRT1 following treatment with Notch1 inhibitor DAPT (20 nM; 24 h). (H) Western blot detection of global lactylation levels. (I) RT-qPCR analysis of microglial polarization markers. (J) Western blot analysis of iNOS and Arg-1 protein expression. (K) Flow cytometric analysis of ROS levels. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; SIRT1, sirtuin 1; RT-qPCR, reverse transcription-quantitative PCR; iNOS, inducible nitric oxide synthase; ROS, reactive oxygen species; NC, negative control; ns, not significant.

Article Snippet: The SIRT1 inhibitor EX527 (10 μ M; 49843-98-3; TargetMol) and the SIRT1 activator SRT1720 (5 μ M; 1001645-58-4; TargetMol) were added separately and incubated for 24 h. The γ-secretase inhibitor DAPT (20 nM; cat. no. HY-13027; MedChemExpress) was administered in parallel experiments, while DMSO served as the vehicle control.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control