Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: PGC1α regulates the mitochondrial metabolism response to cyclic stretch, which inhibits neointimal hyperplasia

doi: 10.1007/s00018-025-05790-x

Figure Lengend Snippet: Physiological cyclic stretch (CS) increased VSMC mitochondrial mass and ATP production in vitro. ( A ) Representative ultrasound images of the carotid artery on the self-control side and injured side at 4 weeks postinjury were used to measure the change in the carotid arterial inner diameter ( n = 4). ( B ) Flow chart depicting the process of multimode ultrasound imaging in the intimal injury model. In an in vitro study, cyclic stretch (CS) mimicked the control group in vivo, and the static condition (ST) mimicked the in vivo injury group. ( C ) Representative immunofluorescence images of the distribution of mitochondria (red) ( n = 16). Blue: DAPI; scale bar = 20 μm. ( D ) mtDNA levels were determined by qRT‒PCR ( n = 6). ( E ) Representative immunofluorescence images of JC-1 staining used to determine the membrane potential of mitochondria ( n = 4). Blue: DAPI; red: JC-1 aggregates; green: JC-1 monomers. Scale bar = 20 μm. ( F ) MFN2 and DRP1 mRNA levels were determined by qRT‒PCR ( n = 4). ( G ) VSMC mRNA levels of key rate-limiting enzymes in the mitochondrial function pathway were determined by qRT‒PCR ( n = 4). ( H ) ATP content was determined by a luciferase assay system ( n = 4). P values were calculated by a two-tailed Mann‒Whitney test. The values are shown as the means ± SD . * P < 0.05

Article Snippet: Primary rat aortic VSMCs were characterized by immunofluorescence staining for the VSMC marker smooth muscle-specific α-actin (SMA) (Proteintech, China), and passages 4 to 7 of the VSMCs and cell populations with greater than 95% purity were used in the present study.

Techniques: In Vitro, Control, Imaging, In Vivo, Immunofluorescence, Staining, Membrane, Luciferase, Two Tailed Test

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: PGC1α regulates the mitochondrial metabolism response to cyclic stretch, which inhibits neointimal hyperplasia

doi: 10.1007/s00018-025-05790-x

Figure Lengend Snippet: The PGC1α activator ZLN005 promoted VSMC mitochondrial function, but siRNA targeting PGC1α inhibited VSMC mitochondrial function. ( A ) The protein level of PGC1α determined by western blot under control and ZLN005 (10 µM) conditions for 48 h ( n = 5). ( B ) The mRNA level of mtDNA after treatment with ZLN005 was determined by qRT‒PCR ( n = 5). ( C ) Representative immunofluorescence images of the distribution of mitochondria (red) in VSMCs after treatment with ZLN005 ( n = 15). Blue: DAPI; scale bar = 20 μm. ( D ) The mRNA levels of MFN2 and DRP1 after treatment with ZLN005 were determined by qRT‒PCR ( n = 4). ( E ) Representative immunofluorescence images of JC-1 staining used to determine the membrane potential of mitochondria after treatment with ZLN005 ( n = 4). Blue: DAPI; red: JC-1 aggregates; green: JC-1 monomers. Scale bar = 20 μm. ( F ) ATP production was determined by a luciferase assay after treatment with ZLN005 ( n = 4). ( G ) The protein level of PGC1α in VSMCs treated with siRNA was determined by western blotting ( n = 5). ( H ) The mRNA level of mtDNA after siRNA treatment was determined by qRT‒PCR ( n = 4). ( I ) Representative immunofluorescence images of the distribution of mitochondria after siRNA treatment ( n = 15). Blue: DAPI; red: MitoTracker Red. Scale bar = 20 μm. ( J ) The mRNA levels of MFN2 and DRP1 in VSMCs treated with siRNAs were determined by qRT‒PCR ( n = 4). ( K ) Representative immunofluorescence images of JC-1 staining used to determine the membrane potential of mitochondria after siRNA treatment ( n = 4). Blue: DAPI; red: JC-1 aggregates; green: JC-1 monomers. Scale bar = 20 μm. () ATP content in VSMCs transfected with siRNA was determined by a luciferase assay system ( n = 4). P values were calculated by a two-tailed Mann‒Whitney test. The values are shown as the means ± SD . * P < 0.05

Article Snippet: Primary rat aortic VSMCs were characterized by immunofluorescence staining for the VSMC marker smooth muscle-specific α-actin (SMA) (Proteintech, China), and passages 4 to 7 of the VSMCs and cell populations with greater than 95% purity were used in the present study.

Techniques: Western Blot, Control, Immunofluorescence, Staining, Membrane, Luciferase, Transfection, Two Tailed Test

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: PGC1α regulates the mitochondrial metabolism response to cyclic stretch, which inhibits neointimal hyperplasia

doi: 10.1007/s00018-025-05790-x

Figure Lengend Snippet: PGC1α knockout repressed mitochondrial function in human aortic VSMCs. ( A ) The ultrastructures of control and PGC1α knockout human aortic VSMC mitochondria were visualized by TEM. Scale bar = 500 nm. ( B ) Representative immunofluorescence images of human aortic VSMC mitochondria labeled with MitoTracker Red acquired by 3D-structured illumination microscopy ( n = 13). Scale bar = 5 μm. (C) PLS-DA score plot of the untargeted metabolomics study. ( D ) Hierarchical clustering heatmaps of differentially abundant metabolites identified via metabolomics analysis between the control group and the PGC1α knockout group. ( E ) Differential enrichment score map of the metabolomics. P values were calculated by a two-tailed Mann‒Whitney test. The values are shown as the means ± SD . * P < 0.05

Article Snippet: Primary rat aortic VSMCs were characterized by immunofluorescence staining for the VSMC marker smooth muscle-specific α-actin (SMA) (Proteintech, China), and passages 4 to 7 of the VSMCs and cell populations with greater than 95% purity were used in the present study.

Techniques: Knock-Out, Control, Immunofluorescence, Labeling, Microscopy, Two Tailed Test

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: PGC1α regulates the mitochondrial metabolism response to cyclic stretch, which inhibits neointimal hyperplasia

doi: 10.1007/s00018-025-05790-x

Figure Lengend Snippet: The PGC1α activator ZLN005 effectively improved mitochondrial function and inhibited VSMC hyperproliferation in vivo. (A) Flow chart of the treatment with ZLN005 in the intimal injury model and the experiments after 4 weeks. (B) The mRNA levels of PGC1α and PGC1β in the carotid artery in the injured group compared with those in the treatment group were determined by qRT‒PCR ( n = 4). (C) The protein level of PGC1α in the carotid artery in the injured group with or without ZLN005 treatment was determined by western blotting. (D) Representative immunofluorescence images of the carotid artery in the injured group and ZLN005 treatment group at 4 weeks ( n ≥ 3). Blue: DAPI; green: SMA; red: PGC1α. Scale bar = 50 μm. (E) The mRNA levels of rate-limiting enzymes in the carotid artery in the injured group compared with those in the treatment group were determined by qRT‒PCR ( n = 4). (F) The mRNA levels of target genes of PGC1α in the carotid artery in the injured group compared with those in the treatment group were determined by qRT‒PCR ( n = 4). (G) Representative HE-stained images of carotid arteries from the injured group and treatment group at 4 weeks ( n = 4). Scale bar = 50 μm. P values were calculated by a two-tailed Mann‒Whitney test. The values are the means ± SD . * P < 0.05

Article Snippet: Primary rat aortic VSMCs were characterized by immunofluorescence staining for the VSMC marker smooth muscle-specific α-actin (SMA) (Proteintech, China), and passages 4 to 7 of the VSMCs and cell populations with greater than 95% purity were used in the present study.

Techniques: In Vivo, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: PGC1α regulates the mitochondrial metabolism response to cyclic stretch, which inhibits neointimal hyperplasia

doi: 10.1007/s00018-025-05790-x

Figure Lengend Snippet: Schematic diagram of PGC1α regulation of VSMC mitochondrial function in vivo and in vitro. (A) In vivo, the intimal injury arteries were harvested after four weeks, and the opposite carotid artery without treatment was collected for the control. Intimal injury caused neointimal hyperplasia and decreased the expression of PGC1α, which consequently suppressed mitochondrial biogenesis, metabolism, and dynamics. However, the subcutaneous injection of ZLN005 every other day reversed these effects and partially restored mitochondrial function. In addition, an abnormal ultrastructure of mitochondria occurred in human plaques, and PGC1α knockout in human aortic VSMCs impaired mitochondrial metabolism. (B) In vitro, cyclic stretch activated Smad3 phosphorylation and then increased the expression levels of PGC1α in VSMCs. PGC1α further activated the transcription of downstream target genes involved in mitochondrial biogenesis, metabolism, and dynamics, ultimately promoting the mitochondrial function of VSMCs

Article Snippet: Primary rat aortic VSMCs were characterized by immunofluorescence staining for the VSMC marker smooth muscle-specific α-actin (SMA) (Proteintech, China), and passages 4 to 7 of the VSMCs and cell populations with greater than 95% purity were used in the present study.

Techniques: In Vivo, In Vitro, Control, Expressing, Injection, Knock-Out, Phospho-proteomics

Journal: Molecular metabolism

Article Title: Hepatic stellate cell-specific Kcnma1 deletion mitigates metabolic dysfunction-associated steatotic liver disease progression via upregulating Amphiregulin secretion.

doi: 10.1016/j.molmet.2025.102164

Figure Lengend Snippet: Figure 3. (A and B) Representative results of F4/80 and αSMA immunofluorescence staining

Article Snippet: For the immunofluorescence staining of the inflammatory marker F4/80 (70076, CST, Rabbit/IgG, 1:800) and liver fibrosis marker αSMA (19245, CST, Rabbit/IgG, 1:1000) in the mouse liver sections, perform the staining operation and imaging analysis according to the method described in the above "Immunofluorescence staining assays".

Techniques: Immunofluorescence, Staining

Journal: Andrology

Article Title: The use of deidentified organ donor testes for research.

doi: 10.1111/andr.70008

Figure Lengend Snippet: FIGURE 5 Utilization of human pre-pubertal and adult testis tissues as antibody controls in hPSC research. (A) The immunostaining of adult (24 y.o.) human testis and human pluripotent stem cell (hPSC)-derived testis-like organoids (Days 26 and 35) for DMRT1 and WT1 markers of supporting cells, steroidogenic cell marker CYP11A1, and peritubular cell marker αSMA (CST, 56856, 1:400), as well as for blood–testis barrier markers ZO1 (CST, 13663, 1:400) and Occludin (CST, 91131, 1:400). (B) The immunostaining of pre-pubertal (11 y.o.) human testis for supporting cell markers WT1, GATA4, SOX9, and AMH. (C) The immunostaining of hPSC-derived testis-like organoids (Days 20 and 16) for supporting cell markers GATA4, SOX9, AMH, and mesonephric marker PAX2. Scale bars 50 µm. All samples and images were prepared as previously described.40

Article Snippet: F IGURE 5 Utilization of human pre-pubertal and adult testis tissues as antibody controls in hPSC research. (A) The immunostaining of adult (24 y.o.) human testis and human pluripotent stem cell (hPSC)-derived testis-like organoids (Days 26 and 35) for DMRT1 andWT1markers of supporting cells, steroidogenic cell marker CYP11A1, and peritubular cell marker αSMA (CST, 56856, 1:400), as well as for blood–testis barrier markers ZO1 (CST, 13663, 1:400) andOccludin (CST, 91131, 1:400). (B) The immunostaining of pre-pubertal (11 y.o.) human testis for supporting cell markersWT1, GATA4, SOX9, and AMH. (C) The immunostaining of hPSC-derived testis-like organoids (Days 20 and 16) for supporting cell markers GATA4, SOX9, AMH, andmesonephric marker PAX2.

Techniques: Immunostaining, Derivative Assay, Marker

Journal: Andrology

Article Title: The use of deidentified organ donor testes for research.

doi: 10.1111/andr.70008

Figure Lengend Snippet: FIGURE 5 Utilization of human pre-pubertal and adult testis tissues as antibody controls in hPSC research. (A) The immunostaining of adult (24 y.o.) human testis and human pluripotent stem cell (hPSC)-derived testis-like organoids (Days 26 and 35) for DMRT1 and WT1 markers of supporting cells, steroidogenic cell marker CYP11A1, and peritubular cell marker αSMA (CST, 56856, 1:400), as well as for blood–testis barrier markers ZO1 (CST, 13663, 1:400) and Occludin (CST, 91131, 1:400). (B) The immunostaining of pre-pubertal (11 y.o.) human testis for supporting cell markers WT1, GATA4, SOX9, and AMH. (C) The immunostaining of hPSC-derived testis-like organoids (Days 20 and 16) for supporting cell markers GATA4, SOX9, AMH, and mesonephric marker PAX2. Scale bars 50 µm. All samples and images were prepared as previously described.40

Article Snippet: F IGURE 5 Utilization of human pre-pubertal and adult testis tissues as antibody controls in hPSC research. (A) The immunostaining of adult (24 y.o.) human testis and human pluripotent stem cell (hPSC)-derived testis-like organoids (Days 26 and 35) for DMRT1 andWT1markers of supporting cells, steroidogenic cell marker CYP11A1, and peritubular cell marker αSMA (CST, 56856, 1:400), as well as for blood–testis barrier markers ZO1 (CST, 13663, 1:400) andOccludin (CST, 91131, 1:400). (B) The immunostaining of pre-pubertal (11 y.o.) human testis for supporting cell markersWT1, GATA4, SOX9, and AMH. (C) The immunostaining of hPSC-derived testis-like organoids (Days 20 and 16) for supporting cell markers GATA4, SOX9, AMH, andmesonephric marker PAX2.

Techniques: Immunostaining, Derivative Assay, Marker