Journal: Journal of translational medicine

Article Title: Nicotinamide mononucleotide combined with PJ-34 protects microglial cells from lipopolysaccharide-induced mitochondrial impairment through NMNAT3-PARP1 axis.

doi: 10.1186/s12967-025-06280-1

Figure Lengend Snippet: Fig. 4 NMN supplementation up-regulates NMNAT3, down-regulates PARP1 and restores cell viability. (a) BV2 cells were treated with different concentra tions of NMN for 24 h and viability was measured with MTT assay (n = 6; **, p < 0.01). (b) BV2 cells were treated with LPS and different concentrations of NMN for 24 h and viability was measured with MTT assay (n = 5; ###, p < 0.001; *, p < 0.05). (c) Representative immunoblots of NMNAT3, PARP1 and PAR poly mer in LPS-induced BV2 cells treated with different concentration of NMN. β-actin was used as a loading control. (d) Quantification of relative expression levels of NMNAT3 protein in each group (n = 3; #, p < 0.05; *, p < 0.05). (e) Quantification of relative expression levels of PARP1 protein in each group (n = 5; #, p < 0.05; **, p < 0.01; ***, p < 0.001). (f) Quantification of total PARylation levels in each group (n = 4; #, p < 0.05, *, p < 0.05). (g) Schematic representation of production and metabolism of NAD+ in the nucleus and mitochondria. Abbreviations: NMN, nicotinamide mononucleotide; NMNAT1, Nicotinamide mononucleotide adenylyl transferase 1; NMNAT3, Nicotinamide mononucleotide adenylyl transferase 3; NAD+, Nicotinamide adenine dinucleotide; NAM, nicotinamide; NAMPT, Nicotinamide phosphoribosyltransferase; PJ-34, 2-(dimethylamino)-N-(5,6-dihydro-6-oxophenanthridin-2yl) acetamide; PARP1, Poly (ADP-ribose) polymerase-1; PAR, poly ADP-ribose. Data are presented as mean ± SEM. Statistical analyses were performed with One-way ANOVA followed by Dunnett’s post hoc test (a, b, d, e, f)

Article Snippet: Microglial activation model was induced by 1 μg/mL LPS [31, 32] (M9524, Abmole, TX, USA) which was followed by different concentration of NMN (S31451, Shanghai Yuanye Bio-Technology, China) or PJ-34 (HY-13688 A, MCE, NJ, USA) treatment.

Techniques: MTT Assay, Western Blot, Concentration Assay, Control, Expressing

Journal: Journal of translational medicine

Article Title: Nicotinamide mononucleotide combined with PJ-34 protects microglial cells from lipopolysaccharide-induced mitochondrial impairment through NMNAT3-PARP1 axis.

doi: 10.1186/s12967-025-06280-1

Figure Lengend Snippet: Fig. 5 NMN supplementation combined with a PARP1 inhibitor jointly restrains PARP1 activity, improves mitochondrial quality. (a) BV2 cells were treated with different concentrations of PJ-34 for 24 h and viability was measured with MTT assay. (n = 6; ***, p < 0.001, ****, p < 0.0001). (b) BV2 cells were treated with NMN and PJ-34 for 24 h and viability was measured with MTT assay (n = 5; #, p < 0.05; ***, p < 0.001; ****, p < 0.0001; ++, p < 0.01). (c) Representative im munoblots of PARP1 and PAR polymer in LPS-induced BV2 cells treated with NMN and PJ-34. β-actin was used as a loading control. (d) Quantification of relative expression levels of PARP1 protein in each group (n = 6; ###, p < 0.001; *, p < 0.05). (e) Quantification of total PARylation levels in each group (n = 6; #, p < 0.5; ***, p < 0.001; ++++, p < 0.0001). (f) Quantification of ATP levels in LPS-induced BV2 cells (n = 7; #p < 0.05; **p < 0.01, ****p < 0.0001). (g) The quantifi cation of the MMP in LPS-induced BV2 cells (n = 3; #p < 0.05; +, p < 0.05). (h) Representative images of JC-1 staining in LPS-induced BV2 cells analyzed by flow cytometry. Data are presented as mean ± SEM. Statistical analyses were performed with One-way ANOVA followed by Dunnett’s post hoc test (a) or Turkey’s post hoc test (b, d, e, f, g). Abbreviations: LPS, lipopolysaccharide; NMN, nicotinamide mononucleotide; PARP1, Poly (ADP-ribose) polymerase-1; PAR, poly ADP-ribose; MMP, Mitochondrial membrane potential

Article Snippet: Microglial activation model was induced by 1 μg/mL LPS [31, 32] (M9524, Abmole, TX, USA) which was followed by different concentration of NMN (S31451, Shanghai Yuanye Bio-Technology, China) or PJ-34 (HY-13688 A, MCE, NJ, USA) treatment.

Techniques: Activity Assay, MTT Assay, Polymer, Control, Expressing, Staining, Flow Cytometry, Membrane

Journal: Journal of translational medicine

Article Title: Nicotinamide mononucleotide combined with PJ-34 protects microglial cells from lipopolysaccharide-induced mitochondrial impairment through NMNAT3-PARP1 axis.

doi: 10.1186/s12967-025-06280-1

Figure Lengend Snippet: Fig. 6 NMN supplementation combined with a PARP1 inhibitor jointly improves mitophagy. (a) Representative immunoblots of mitophagy-related proteins in LPS-induced BV2 cells treated with NMN and PJ-34. GAPDH and β-actin was used as a loading control. (b) Quantification of relative expres sion levels of Beclin1 protein in each group (n = 6; ##, p < 0.01; ***, p < 0.001; ****, p < 0.0001). (c) Quantification of relative expression levels of LC3II/LC3I in each group (n = 6; #, p < 0.05; ***, p < 0.001; ++, p < 0.01). (d) Quantification of relative expression levels of p62 protein in each group (n = 6; #, p < 0.05; ***, p < 0.001; ****, p < 0.0001). (e) Quantification of relative expression levels of PINK1 protein in each group (n = 4; ##, p < 0.01; *, p < 0.05). (f) Quantification of relative expression levels of Parkin protein in each group (n = 6; #, p < 0.05; ***, p < 0.001; +, p < 0.05). (g) Pearson’s r value of the colocalization of LAMP1 and mitochondria in LPS-induced BV2 cells (n = 6; ####, p < 0.0001; *, p < 0.05; ****, p < 0.0001; ++, p < 0.01). (h) Representative images of colocalization of LAMP1 (green) and mito-tracker (red) in LPS-induced BV2 cells. Scale bar = 10 μm. Data are presented as mean ± SEM. Statistical analyses were performed with One-way ANOVA followed by Turkey’s post hoc test (b-g). Abbreviations: LPS, lipopolysaccharide; NMN, nicotinamide mononucleotide; PINK1, PTEN induced kinase 1; LAMP1, Lysosomal-associated membrane protein 1

Article Snippet: Microglial activation model was induced by 1 μg/mL LPS [31, 32] (M9524, Abmole, TX, USA) which was followed by different concentration of NMN (S31451, Shanghai Yuanye Bio-Technology, China) or PJ-34 (HY-13688 A, MCE, NJ, USA) treatment.

Techniques: Western Blot, Control, Expressing, Membrane

Journal: Journal of translational medicine

Article Title: Nicotinamide mononucleotide combined with PJ-34 protects microglial cells from lipopolysaccharide-induced mitochondrial impairment through NMNAT3-PARP1 axis.

doi: 10.1186/s12967-025-06280-1

Figure Lengend Snippet: Fig. 7 NMN supplementation combined with a PARP1 inhibitor jointly improves mitochondrial dynamics and mitochondrial morphology. (a) Repre sentative immunoblots of mitochondrial fission and fusion proteins in LPS-induced BV2 cells treated with NMN and PJ-34. β-actin was used as a loading control. (b) Quantification of relative expression levels of p-Drp1(Ser616) protein in each group (n = 4; #, p < 0.05; **, p < 0.01; +, p < 0.05). (c) Quantification of relative expression levels of Mfn2 protein in each group (n = 4; #, p < 0.05; **, p < 0.01; ****, p < 0.0001; ++++, p < 0.0001). (d) Quantification of relative ex pression levels of Opa1 protein in each group (n = 4; ##, p < 0.01; **, p < 0.01; ****, p < 0.0001). Data are presented as mean ± SEM. (e) Quantitation analysis of mitochondrial length in each group (n = 4; ####, p < 0.0001; **, p < 0.01; ****, p < 0.0001) (f) The Representative TEM images of LPS-induced BV2 cells treated with NMN and PJ-34. Scale bar: upper, 500 nm; lower, 1 μm. Data are presented as mean ± SEM. Statistical analyses were performed with One-way ANOVA followed by Turkey’s post hoc test (b-e). Abbreviations: LPS, lipopolysaccharide; NMN, nicotinamide mononucleotide

Article Snippet: Microglial activation model was induced by 1 μg/mL LPS [31, 32] (M9524, Abmole, TX, USA) which was followed by different concentration of NMN (S31451, Shanghai Yuanye Bio-Technology, China) or PJ-34 (HY-13688 A, MCE, NJ, USA) treatment.

Techniques: Western Blot, Control, Expressing, Quantitation Assay

Journal: Journal of translational medicine

Article Title: Nicotinamide mononucleotide combined with PJ-34 protects microglial cells from lipopolysaccharide-induced mitochondrial impairment through NMNAT3-PARP1 axis.

doi: 10.1186/s12967-025-06280-1

Figure Lengend Snippet: Fig. 8 PARP1 inhibition enhances PINK1/Parkin-mediated mitophagy by increasing the binding of FoxO1 to the PINK1 promoter. (a) The effects of PARP1 on PINK1 mRNA expression of BV2 cells were detected by RT-qPCR (n = 3; #, p < 0.05; ***, p < 0.001; ++, p < 0.01). (b) The effects of PARP1 on FoxO1 binding to PINK1 promoter of BV2 cells were detected by ChIP (n = 3; ##, p < 0.01; ****, p < 0.0001, ++++, p < 0.0001). (c) Schematic representation of PARP1 regulates the binding of FoxO1 to PINK1 promoter. (d) Schematic representation of the effect of LPS on the binding of FoxO1 to PINK1 promoter. (e) Schematic representation of the effect of NMN combined with PJ-34 on the binding of FoxO1 to PINK1 promoter. Abbreviations: LPS, lipopolysaccharide; NMN, nicotinamide mononucleotide; NMNAT1, Nicotinamide mononucleotide adenylyl transferase 1; NMNAT3, Nicotinamide mononucleotide adenylyl trans ferase 3; NAD+, Nicotinamide adenine dinucleotide; PARP1, Poly (ADP-ribose) polymerase-1; FoxO1, Forkhead box O1. Data are presented as mean ± SEM. Statistical analyses were performed with One-way ANOVA followed by Turkey’s post hoc test (a) or Two-way ANOVA followed by Turkey’s post hoc test (b)

Article Snippet: Microglial activation model was induced by 1 μg/mL LPS [31, 32] (M9524, Abmole, TX, USA) which was followed by different concentration of NMN (S31451, Shanghai Yuanye Bio-Technology, China) or PJ-34 (HY-13688 A, MCE, NJ, USA) treatment.

Techniques: Inhibition, Binding Assay, Expressing, Quantitative RT-PCR

Journal: eLife

Article Title: Large-scale characterization of drug mechanism of action using proteome-wide thermal shift assays

doi: 10.7554/eLife.95595

Figure Lengend Snippet:

Article Snippet: Chemical compound , PJ34 (hydrochloride) , MedChemExpress , HY-13688 , .

Techniques: Recombinant, Software

Journal: International Journal of Molecular Sciences

Article Title: PARP14 Contributes to the Development of the Tumor-Associated Macrophage Phenotype

doi: 10.3390/ijms25073601

Figure Lengend Snippet: PARP14 mediated M2 polarization in BMDMs. Bone-marrow-derived macrophages (BMDMs) from C57/BL6 mice were differentiated with 25 ng/mL M-CSF for 6 days. BMDMs were pre-treated with either 20 μM PJ34 ( A ) or 50 μM MCD113 ( B ) for 1 h and then polarized into M2 macrophages with 5 ng/mL IL-4 for 24 h. On day 7, mRNA levels of the M2 markers Arg1 , Fizz1 , and MRC1 were determined by RT-qPCR. BMDMs were also differentiated from PARP14 KO mice and their wild type counterparts with 25 ng/mL M-CSF for 6 days and then polarized into M2 with 5 ng/mL IL-4 for 24 h. RNA extraction was performed on day 7 followed by RT-qPCR ( C ). Light blue and purple colors in ( C ) represent WT and PARP14 KO groups, respectively. The graphs show the mean of at least 3 independent experiments (±SEM). Statistical evaluation was performed with the Kruskal–Wallis test followed by Dunn’s post-hoc test ( A ) ( Arg1 in ( B )) or with one-way ANOVA and Tukey’s post-hoc test ( C ) ( Fizz1 and MRC1 in ( B )) (* p < 0.05).

Article Snippet: PJ34 (2-(dimethylamino)-N-(6-oxo-5H-phenanthridin-2-yl)acetamide hydrochloride, 344458-15-7) was obtained from Sigma-Aldrich (Budapest, Hungary), and MCD113 was synthesized by Dana Ferraris’s group at McDaniel College (Westminster, MD, USA), as previously reported [ ]. (In the cited reference, MCD113 was referred to as compound 4t, and the IC 50 of the inhibitor was 160 nM for PARP14.)

Techniques: Derivative Assay, Quantitative RT-PCR, RNA Extraction

Journal: International Journal of Molecular Sciences

Article Title: PARP14 Contributes to the Development of the Tumor-Associated Macrophage Phenotype

doi: 10.3390/ijms25073601

Figure Lengend Snippet: Soluble factors produced by 4T1 murine breast cancer cells reprogrammed murine BMDMs toward an M2-like TAM phenotype in a PARP14-dependent manner. Bone-marrow-derived macrophages (BMDMs) were differentiated from C57/BL6 mice with 25 ng/mL M-CSF for 6 days. Cells were pre-treated with either 20 μM PJ34 ( A ) or 50 μM MCD113 ( B ) for 1 h and then treated with 4T1 conditioned media (4T1CM; diluted at a 1:1 ratio with full DMEM) for 24 h to polarize them into TAMs. On day 7, mRNA levels of M2 markers were quantified by RT-qPCR. BMDMs were also differentiated from PARP14 KO mice and their wild-type counterparts with 25 ng/mL M-CSF for 6 days and then treated with 4T1CM for 24 h. RNA extraction was performed on day 7 followed by RT-qPCR ( C ). Dark blue and grey colors in ( C ) represent WT and PARP14 KO groups, respectively. The graphs show the mean of at least 3 independent experiments (±SEM). Data were analyzed using the Kruskal–Wallis test and Dunn’s post-hoc test ( A , B ) and with one-way ANOVA followed by Tukey’s post-hoc test ( C ) (* p < 0.05).

Article Snippet: PJ34 (2-(dimethylamino)-N-(6-oxo-5H-phenanthridin-2-yl)acetamide hydrochloride, 344458-15-7) was obtained from Sigma-Aldrich (Budapest, Hungary), and MCD113 was synthesized by Dana Ferraris’s group at McDaniel College (Westminster, MD, USA), as previously reported [ ]. (In the cited reference, MCD113 was referred to as compound 4t, and the IC 50 of the inhibitor was 160 nM for PARP14.)

Techniques: Produced, Derivative Assay, Quantitative RT-PCR, RNA Extraction

Journal: International Journal of Molecular Sciences

Article Title: PARP14 Contributes to the Development of the Tumor-Associated Macrophage Phenotype

doi: 10.3390/ijms25073601

Figure Lengend Snippet: The PARP inhibitor PJ34 and the PARP14 inhibitor MCD113 reduced the expression of M2 markers in human macrophages. Macrophages were differentiated from human monocytes with 50 ng/mL M-CSF for 5 days. On day 5, they were pre-treated with either 20 μM PJ34 or 50 μM MCD113 for 1 h and then stimulated with 100 ng/mL IL-4 for 24 h to polarize them into M2 macrophages. On day 6, cells were collected for RNA extraction, and M2 markers were measured with RT-qPCR. Histograms show the mean of at least 3 independent experiments (±SEM). Statistical evaluation was performed with the Kruskal–Wallis test and Dunn’s post-hoc test (* p < 0.05) ( A ). On day 6, cells were also collected and labelled with APC-conjugated anti-human mouse CD206 monoclonal antibody. Fluorescence intensities and the ratio of CD206-expressing cells were measured by flow cytometry. The mean value of MFIs (median fluorescence intensities) of the cell surface molecule CD206 was calculated from 8 independent experiments (±SEM). The mean value of frequency of CD206+ cells was calculated from 5 independent experiments (±SEM). The relative frequency of CD206-expressing cells was determined as follows. For each donor, the frequency of CD206+ cells differentiated in the presence of IL-4 was considered to be 1, and the frequency of untreated cells and cells treated with IL-4 + PARP inhibitors was compared to this. Data were analyzed by one-way ANOVA followed by Bonferroni’s post-hoc test (left panel) or by the Kruskal–Wallis test and Dunn’s post-hoc test (middle and right panels) (* p < 0.05) ( B ).

Article Snippet: PJ34 (2-(dimethylamino)-N-(6-oxo-5H-phenanthridin-2-yl)acetamide hydrochloride, 344458-15-7) was obtained from Sigma-Aldrich (Budapest, Hungary), and MCD113 was synthesized by Dana Ferraris’s group at McDaniel College (Westminster, MD, USA), as previously reported [ ]. (In the cited reference, MCD113 was referred to as compound 4t, and the IC 50 of the inhibitor was 160 nM for PARP14.)

Techniques: Expressing, RNA Extraction, Quantitative RT-PCR, Fluorescence, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: PARP14 Contributes to the Development of the Tumor-Associated Macrophage Phenotype

doi: 10.3390/ijms25073601

Figure Lengend Snippet: The PARPi PJ34 and the PARP14i MCD113 decreased M2 marker gene expression in JIMT-1-THP-1-derived MΦ co-culture spheroids. Spheroids were grown from co-cultures of JIMT-1 cells and THP-1-derived MΦs or from THP-1-derived MΦs only, as described in . Spheroids were kept in cultures for 4 days after formation (1 day in the presence of 20 ng/mL IFN-γ), followed by treatments with either 25 μM PJ34 ( A ) or 55 μM MCD113 ( B ) for 24 h. Columns show the relative expression of mRNA levels of M2 markers in spheroids and spheroid co-cultures quantified by RT-qPCR. The histograms represent the mean of at least 3 independent experiments (±SEM). Statistical evaluation was performed with one-way ANOVA and Tukey’s post-hoc test ( MRC1 and CCL17 in ( A ), Fibronectin in ( A , B )), or with the Kruskal–Wallis test and Dunn’s post-hoc test (* p < 0.05, ns: not significant).

Article Snippet: PJ34 (2-(dimethylamino)-N-(6-oxo-5H-phenanthridin-2-yl)acetamide hydrochloride, 344458-15-7) was obtained from Sigma-Aldrich (Budapest, Hungary), and MCD113 was synthesized by Dana Ferraris’s group at McDaniel College (Westminster, MD, USA), as previously reported [ ]. (In the cited reference, MCD113 was referred to as compound 4t, and the IC 50 of the inhibitor was 160 nM for PARP14.)

Techniques: Marker, Expressing, Derivative Assay, Co-Culture Assay, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: PARP14 Contributes to the Development of the Tumor-Associated Macrophage Phenotype

doi: 10.3390/ijms25073601

Figure Lengend Snippet: The PARP inhibitors PJ34 and MCD113 induced apoptotic tumor cell death in spheroid co-cultures. Co-culture spheroids were generated with JIMT-1-EGFP (green) and Cell Tracker Blue-stained derived MΦs at a 1:1 ratio. Spheroids were kept in cultures for a duration of 4 days (1 day in the presence of 20 ng/mL IFN-γ) and were then treated with 25 μM PJ34 or 55 μM MCD113 for 24 h. Cells were stained with Annexin V 647 to detect apoptosis. Images were taken after spheroid formation on day 5 following PJ34 and MCD113 treatments (( A , B ) respectively). Images of 3 spheroids/treatment were taken and analyzed for the fluorescence intensity of Annexin V-Alexa 647 in tumor cells/each well ( C , D ). Means ± SEM of 3 independent experiments are shown. The statistics were calculated with one-way ANOVA followed by Dunnett’s post-hoc test (* p < 0.05).

Article Snippet: PJ34 (2-(dimethylamino)-N-(6-oxo-5H-phenanthridin-2-yl)acetamide hydrochloride, 344458-15-7) was obtained from Sigma-Aldrich (Budapest, Hungary), and MCD113 was synthesized by Dana Ferraris’s group at McDaniel College (Westminster, MD, USA), as previously reported [ ]. (In the cited reference, MCD113 was referred to as compound 4t, and the IC 50 of the inhibitor was 160 nM for PARP14.)

Techniques: Co-Culture Assay, Generated, Staining, Derivative Assay, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: PARP14 Contributes to the Development of the Tumor-Associated Macrophage Phenotype

doi: 10.3390/ijms25073601

Figure Lengend Snippet: Proteome profiling suggesting the involvement of LCN2, MIF, and PAI-1 in TAM polarization. Spheroids were generated either from THP-1-derived MΦs or JIMT-1-THP-1-MΦ co-cultures. Cells were kept in cultures for 4 days (1 day in the presence of 20 ng/mL IFN-γ), and supernatant was collected. Cell supernatants (500 μL) were added to each membrane array ( A ), and analytes were detected by proteome profiling as described in ( B ). Using the same conditions, RNA was also extracted from spheroids, and mRNA levels of the same set of cytokines were quantified by RT-qPCR. Treatments with the PARPi PJ34 (25 μM) and PARP14i MCD113 (55 μM) resulted in a significant decrease in the expression of these cytokines ( C ). The numbers in ( A , B ) indicate the different cytokines detected. Red rectangles in ( B ) depict the three cytokines selected for quantitation by RT-qPCR, as shown in ( C ). Graphs show mean of at least 3 independent experiments (±SEM). Statistical evaluation was performed with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

Article Snippet: PJ34 (2-(dimethylamino)-N-(6-oxo-5H-phenanthridin-2-yl)acetamide hydrochloride, 344458-15-7) was obtained from Sigma-Aldrich (Budapest, Hungary), and MCD113 was synthesized by Dana Ferraris’s group at McDaniel College (Westminster, MD, USA), as previously reported [ ]. (In the cited reference, MCD113 was referred to as compound 4t, and the IC 50 of the inhibitor was 160 nM for PARP14.)

Techniques: Generated, Derivative Assay, Membrane, Quantitative RT-PCR, Expressing, Quantitation Assay

Journal: Animal cells and systems

Article Title: Restoration of NAD + homeostasis protects C2C12 myoblasts and mouse levator ani muscle from mechanical stress-induced damage.

doi: 10.1080/19768354.2022.2106303

Figure Lengend Snippet: Figure 2. Inhibition of PARP-1 activity rescues cell damage induced by MS by increasing NAD+ levels and improving mitochondrial dys- function. To inhibit PARP-1, C2C12 cells were treated with vehicle (PBS) or 1 μM PJ34 hydrochloride for 24 h before applying MS. (A) NAD+

Article Snippet: To evaluate the effect of NMN supplementation, C2C12 cells were treated with vehicle (PBS) or 500 μM NMN (MACKLIN, Shanghai, China) for 24 h after applying MS. To inhibit PARP-1, C2C12 cells were treated with vehicle (PBS) or 1 μM PJ34 hydrochloride (MedChemExpress, Monmouth Junction, NJ, USA) for 24 h before MS.

Techniques: Inhibition, Activity Assay

Journal: Animal cells and systems

Article Title: Restoration of NAD + homeostasis protects C2C12 myoblasts and mouse levator ani muscle from mechanical stress-induced damage.

doi: 10.1080/19768354.2022.2106303

Figure Lengend Snippet: Figure 3. NMN supplementation rescues cell damage induced by mechanical overload by increasing NAD+ levels and improving mito- chondrial dysfunction. To evaluate the effect of NMN supplementation, C2C12 cells were treated with vehicle (PBS) or 500 μM NMN for 24 h before applying MS. (A) NAD+ levels in C2C12 cells detected using a commercial kit. (B and C) ATP content (B) and mitochondrial membrane potential (ΔΨm, C) were measured to assess mitochondrial function. (D) Cell viability was detected using the Cell Counting Kit-8 (CCK8) kit. All experiments were independently repeated at least three times. Statistical data are expressed as the mean ± stan- dard deviation. Differences between the two groups were determined using Dunnett’s t-test. Tukey’s test was performed for multiple comparisons (compared to the control group: ***p < 0.001, **p < 0.01; *p < 0.05; ns, not significant; compared to the MS group: ###p < 0.001, ##p < 0.01; #p < 0.05).

Article Snippet: To evaluate the effect of NMN supplementation, C2C12 cells were treated with vehicle (PBS) or 500 μM NMN (MACKLIN, Shanghai, China) for 24 h after applying MS. To inhibit PARP-1, C2C12 cells were treated with vehicle (PBS) or 1 μM PJ34 hydrochloride (MedChemExpress, Monmouth Junction, NJ, USA) for 24 h before MS.

Techniques: Membrane, Cell Counting, Control

Journal: Animal cells and systems

Article Title: Restoration of NAD + homeostasis protects C2C12 myoblasts and mouse levator ani muscle from mechanical stress-induced damage.

doi: 10.1080/19768354.2022.2106303

Figure Lengend Snippet: Figure 6. NMN supplementation rescues LAM damage induced by MS by increasing NAD+ levels and improving mitochondrial dys- function.To evaluate the effect of NMN supplementation, the mice were intraperitoneally injected with vehicle (PBS) or 500 mg NMN/ kg body weight for 7 consecutive days after undergoing VDT. (A) NAD+ levels in the LAM detected using a commercial kit. (B and C) ATP content (B) and mitochondrial membrane potential (ΔΨm, C) were measured to assess mitochondrial function. (D and E) LAM damage was detected using tissue TUNEL staining. The positive staining area was semi-quantified using ImageJ software. All exper- iments were independently repeated at least three times. Statistical data are expressed as the mean ± standard deviation. Differences between the two groups were determined using Dunnett’s t-test. Tukey’s test was performed for multiple comparisons (compared to the control group: ***p < 0.001, **p < 0.01; *p < 0.05; ns, not significant; compared to the VDT group: ###p < 0.001, ##p < 0.01).

Article Snippet: To evaluate the effect of NMN supplementation, C2C12 cells were treated with vehicle (PBS) or 500 μM NMN (MACKLIN, Shanghai, China) for 24 h after applying MS. To inhibit PARP-1, C2C12 cells were treated with vehicle (PBS) or 1 μM PJ34 hydrochloride (MedChemExpress, Monmouth Junction, NJ, USA) for 24 h before MS.

Techniques: Injection, Membrane, TUNEL Assay, Staining, Software, Standard Deviation, Control