Review

gvi ipla2 (Santa Cruz Biotechnology)

Name: gvi ipla2
Brand: Santa Cruz Biotechnology
Rating: 94 (24 reviews)

Santa Cruz Biotechnology is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Santa Cruz Biotechnology gvi ipla2
Gvi Ipla2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gvi ipla2/product/Santa Cruz Biotechnology
Average 94 stars, based on 24 article reviews

gvi ipla2 - by Bioz Stars, 2026-03

94/100 stars

Images

Image Search Results

SLC16A10 modulates T 3 uptake and inflammatory responses in HaCaT cells. A) RT‐qPCR analysis of SLC16A10 mRNA expression in knockdown cells, n = 4; B) Representative western blot showing SLC16A10 protein levels in HaCaT cells transfected with sh‐NC, sh‐SLC16A10‐1, sh‐SLC16A10‐2, or sh‐SLC16A10‐3. GAPDH served as loading control; C) Quantification of SLC16A10 protein expression relative to GAPDH in knockdown cells, n = 3; D) RT‐qPCR analysis of SLC16A10 mRNA expression in overexpression cells, n = 4; E) Representative western blot showing SLC16A10 protein levels in OE‐NC and OE‐SLC16A10 cells; F) Quantification of SLC16A10 protein expression in overexpression cells, n = 4; G) Time‐course analysis of 125 I‐T 3 uptake in sh‐NC and sh‐SLC16A10‐2 cells, n = 3; H) Time‐course analysis of 125 I‐T 3 uptake in OE‐NC and OE‐SLC16A10 cells. Radioactivity measured at indicated time points and expressed as counts per minute (cpm), n = 3; I) Representative western blot showing PLA2, COX‐2, and NF‐κB protein levels in T 3 ‐treated sh‐NC and sh‐SLC16A10‐2 cells; J–L) Quantification of PLA2, COX‐2, and NF‐κB protein expression relative to GAPDH in knockdown cells, n = 3; M) Representative western blot showing inflammatory markers in T 3 ‐treated OE‐NC and OE‐SLC16A10 cells; N–P) Quantification of PLA2, COX‐2, and NF‐κB protein expression in overexpression cells, n = 3; Q–T) ELISA analysis of IL‐6 and TNF‐α protein concentrations in culture supernatants from knockdown and overexpression cells treated with T 3 . * P > 0.05, compared with Control group; & P > 0.05, compared with sh‐NC group; # P > 0.05, compared with OE‐NC group.

Journal: Advanced Science

Article Title: Role of SLC16A10 in Psoriasis Through the Regulation of Arachidonic Acid Metabolism in Keratinocytes

doi: 10.1002/advs.202417093

Figure Lengend Snippet: SLC16A10 modulates T 3 uptake and inflammatory responses in HaCaT cells. A) RT‐qPCR analysis of SLC16A10 mRNA expression in knockdown cells, n = 4; B) Representative western blot showing SLC16A10 protein levels in HaCaT cells transfected with sh‐NC, sh‐SLC16A10‐1, sh‐SLC16A10‐2, or sh‐SLC16A10‐3. GAPDH served as loading control; C) Quantification of SLC16A10 protein expression relative to GAPDH in knockdown cells, n = 3; D) RT‐qPCR analysis of SLC16A10 mRNA expression in overexpression cells, n = 4; E) Representative western blot showing SLC16A10 protein levels in OE‐NC and OE‐SLC16A10 cells; F) Quantification of SLC16A10 protein expression in overexpression cells, n = 4; G) Time‐course analysis of 125 I‐T 3 uptake in sh‐NC and sh‐SLC16A10‐2 cells, n = 3; H) Time‐course analysis of 125 I‐T 3 uptake in OE‐NC and OE‐SLC16A10 cells. Radioactivity measured at indicated time points and expressed as counts per minute (cpm), n = 3; I) Representative western blot showing PLA2, COX‐2, and NF‐κB protein levels in T 3 ‐treated sh‐NC and sh‐SLC16A10‐2 cells; J–L) Quantification of PLA2, COX‐2, and NF‐κB protein expression relative to GAPDH in knockdown cells, n = 3; M) Representative western blot showing inflammatory markers in T 3 ‐treated OE‐NC and OE‐SLC16A10 cells; N–P) Quantification of PLA2, COX‐2, and NF‐κB protein expression in overexpression cells, n = 3; Q–T) ELISA analysis of IL‐6 and TNF‐α protein concentrations in culture supernatants from knockdown and overexpression cells treated with T 3 . * P > 0.05, compared with Control group; & P > 0.05, compared with sh‐NC group; # P > 0.05, compared with OE‐NC group.

Article Snippet: For western blotting (WB) analysis, protein samples from cells or tissues were separated using SDS‐PAGE on 10%–12% gels, transferred onto polyvinylidene fluoride (PVDF) membranes, and probed with primary antibodies against the following proteins: SLC16A10 (1:1000, cat. PA5‐50760; Invitrogen, Carlsbad, CA, USA), β‐actin (1:5000, cat.4 970S; CST), PLA2 (1:500, cat. 22030‐1‐AP; Proteintech, Rosemont, IL, USA), COX‐2 (1:1000, 12375‐1‐AP, Proteintech, Rosemont, IL, USA) and NF‐κB (1:1000, 10745‐1‐AP, Proteintech, Rosemont, IL, USA).

Techniques: Quantitative RT-PCR, Expressing, Knockdown, Western Blot, Transfection, Control, Over Expression, Radioactivity, Enzyme-linked Immunosorbent Assay

MeCP2 lactylation regulates neuronal apoptosis after stroke. A) Heatmap depicting differential lactylation at specific lysine residues of proteins involved in the regulation of neuronal death after stroke. B) PRM quantification of MeCP2 K210 lactylation levels in the brains of sham and MCAO mice ( n = 3 mice per group). C) MeCP2 was immunoprecipitated from sham and MCAO brain tissues, and pan‐Kla and MeCP2 levels were analyzed by Western blot. D) Representative immunofluorescence images showing the proximity ligation assay used to validate MeCP2 lactylation in neurons within the cortex of MCAO mice. Red, in situ PLA detection of MeCP2 and pan‐Kla; green, NeuN; blue, DAPI. E) Heatmap showing differentially lactylated proteins involved in transcriptional regulation between sham and MCAO groups. F) The binding density of MeCP2 was visualized by deepTools: the heatmap presents the CUT&Tag counts at different MeCP2 binding peaks across different treatment groups, ordered by signal strength. G) Distribution of MeCP2 binding peaks across genomic regions in cortical tissues from sham‐operated mice and MCAO mice treated with saline, 2‐DG or 4‐CIN. H) GO enrichment analysis of downregulated and upregulated genes associated with MeCP2 binding sites in the penumbra from MCAO mice. I) Genome browser tracks of MeCP2 CUT&Tag peaks at Pdcd4 and Pla2g6 loci across treatment groups. The purple rectangles indicate the peak regions of MeCP2 on target‐gene promoters. J) Luciferase reporter assay showing relative activity at Pdcd4 and Pla2g6 peaks in HEK293T cells expressing HA‐MeCP2 or HA‐GFP ( n = 3 per group). K) Western blot showing over‐expression of MeCP2 in HEK293 cells, with β‐actin as a loading control. L) Relative luciferase activity at Pdcd4 and Pla2g6 peaks in wild‐type (WT) and MeCP2 −/− HEK293T cells ( n = 4 per group). M) Motif enrichment analysis of MeCP2 binding sites, highlighting top enriched motifs with corresponding p ‐values. N) EMSA demonstrating MeCP2 binding to the CpG‐rich promoters of the target genes Pdcd4 and Pla2g6 . O) ChIP‐qPCR quantification of MeCP2 enrichment at Pdcd4 and Pla2g6 promoter in sham mice and MCAO mice treated with saline, 2‐DG or 4‐CIN ( n = 4 per group). P) Cortical expression of Pdcd4 and Pla2g6 determined by qPCR in sham mice and MCAO mice treated with saline, 2‐DG or 4‐CIN ( n = 5–6 per group). Data are presented as mean ± SEM. # p < 0.05 versus sham control; * p < 0.05, ** p < 0.01, *** p < 0.001 versus MCAO control; ns, not significant.

Journal: Advanced Science

Article Title: MeCP2 Lactylation Protects against Ischemic Brain Injury by Transcriptionally Regulating Neuronal Apoptosis

doi: 10.1002/advs.202415309

Figure Lengend Snippet: MeCP2 lactylation regulates neuronal apoptosis after stroke. A) Heatmap depicting differential lactylation at specific lysine residues of proteins involved in the regulation of neuronal death after stroke. B) PRM quantification of MeCP2 K210 lactylation levels in the brains of sham and MCAO mice ( n = 3 mice per group). C) MeCP2 was immunoprecipitated from sham and MCAO brain tissues, and pan‐Kla and MeCP2 levels were analyzed by Western blot. D) Representative immunofluorescence images showing the proximity ligation assay used to validate MeCP2 lactylation in neurons within the cortex of MCAO mice. Red, in situ PLA detection of MeCP2 and pan‐Kla; green, NeuN; blue, DAPI. E) Heatmap showing differentially lactylated proteins involved in transcriptional regulation between sham and MCAO groups. F) The binding density of MeCP2 was visualized by deepTools: the heatmap presents the CUT&Tag counts at different MeCP2 binding peaks across different treatment groups, ordered by signal strength. G) Distribution of MeCP2 binding peaks across genomic regions in cortical tissues from sham‐operated mice and MCAO mice treated with saline, 2‐DG or 4‐CIN. H) GO enrichment analysis of downregulated and upregulated genes associated with MeCP2 binding sites in the penumbra from MCAO mice. I) Genome browser tracks of MeCP2 CUT&Tag peaks at Pdcd4 and Pla2g6 loci across treatment groups. The purple rectangles indicate the peak regions of MeCP2 on target‐gene promoters. J) Luciferase reporter assay showing relative activity at Pdcd4 and Pla2g6 peaks in HEK293T cells expressing HA‐MeCP2 or HA‐GFP ( n = 3 per group). K) Western blot showing over‐expression of MeCP2 in HEK293 cells, with β‐actin as a loading control. L) Relative luciferase activity at Pdcd4 and Pla2g6 peaks in wild‐type (WT) and MeCP2 −/− HEK293T cells ( n = 4 per group). M) Motif enrichment analysis of MeCP2 binding sites, highlighting top enriched motifs with corresponding p ‐values. N) EMSA demonstrating MeCP2 binding to the CpG‐rich promoters of the target genes Pdcd4 and Pla2g6 . O) ChIP‐qPCR quantification of MeCP2 enrichment at Pdcd4 and Pla2g6 promoter in sham mice and MCAO mice treated with saline, 2‐DG or 4‐CIN ( n = 4 per group). P) Cortical expression of Pdcd4 and Pla2g6 determined by qPCR in sham mice and MCAO mice treated with saline, 2‐DG or 4‐CIN ( n = 5–6 per group). Data are presented as mean ± SEM. # p < 0.05 versus sham control; * p < 0.05, ** p < 0.01, *** p < 0.001 versus MCAO control; ns, not significant.

Article Snippet: The following primary antibodies were used: Anti‐L‐Lactyl Lysine Rabbit mAb (Cat. PTM‐1401, PTM BIO, China), Anti‐lacty‐MeCP2(Lys210) rabbit pAb (PTM BIO, China), Anti‐lacty‐MeCP2(Lys249) rabbit pAb (PTM BIO, China), Rabbit monoclonal anti‐MeCP2 (Cat. 3456, Cell Signaling Technology, USA), Rabbit monoclonal anti‐PDCD4 (Cat. 9535, Cell Signaling Technology, USA), Rabbit monoclonal anti‐HA‐Tag (Cat. 3724, Cell Signaling Technology, USA), Rabbit polyclonal anti‐Cleaved Caspase‐3 (Cat. 9661, Cell Signaling Technology, USA), Rabbit monoclonal anti‐HDAC1 (Cat. 34589, Cell Signaling Technology, USA), Mouse monoclonal anti‐HDAC2 (Cat. 5113, Cell Signaling Technology, USA), Mouse monoclonal anti‐HDAC3 (Cat. 3949, Cell Signaling Technology, USA), Rabbit monoclonal anti‐CHOP (Cat. 5554, Cell Signaling Technology, USA), Rabbit monoclonal anti‐Caspase‐1 (Cat. 24232, Cell Signaling Technology, USA), Rabbit monoclonal anti‐Phospho‐RIP3 (Thr231/Ser232) (Cat. 91702, Cell Signaling Technology, USA), Mouse monoclonal anti‐AlaRS (Cat. sc‐165990, Santa Cruz Biotechnology, USA), Mouse monoclonal anti‐p300 (Cat. sc‐48343, Santa Cruz Biotechnology, USA), Mouse monoclonal anti‐CBP (Cat. sc‐7300, Santa Cruz Biotechnology, USA), Mouse monoclonal anti‐group VI iPLA2 (Cat. sc‐376563, Santa Cruz Biotechnology, USA), Mouse monoclonal anti‐PCAF (Cat. sc‐13124, Santa Cruz Biotechnology, USA), Mouse monoclonal anti‐KAT2A/GCN5(Cat. 66575‐1‐lg, Proteintech, China), Rabbit polyclonal anti‐RIP3 (Cat. 17563‐1‐lg, Proteintech, China), Mouse monoclonal anti‐b‐actin (Cat. 66009‐1‐lg, Proteintech, China), Mouse GAPDH Monoclonal antibodies (Cat. 60004‐1‐Ig, Protsintech, China).

Techniques: Immunoprecipitation, Western Blot, Immunofluorescence, Proximity Ligation Assay, In Situ, Binding Assay, Saline, Luciferase, Reporter Assay, Activity Assay, Expressing, Over Expression, Control, ChIP-qPCR

Lactylation of MeCP2 at K210 and K249 regulates apoptotic gene expression in stroke. A) Sequence alignment of the transcriptional repression domain of MeCP2 across species, highlighting conserved MeCP2 lactylation sites. B) Co‐immunoprecipitation and Western blot analysis showing reduced MeCP2 lactylation levels upon K210R or K249R mutation. Luciferase reporter assays showing increased transcriptional activity at the C) Pdcd4 and D) Pla2g6 promoters in HEK293T cells expressing K210R or K249R MeCP2 mutants compared to wild‐type MeCP2 ( n = 3 per group). E) MeCP2 was immunoprecipitated with anti‐HA from HEK293T cells expressing WT or K210R/K249R mutants, and pan‐Kla and HA‐MeCP2 levels were detected by Western blot. Luciferase reporter assays showing transcriptional activity at the F) Pdcd4 and G) Pla2g6 promoters in HEK293T cells expressing WT MeCP2 or K210/K249 mutants ( n = 4 per group). H) EMSA analysis showing recombinant wild‐type and mutant MeCP2 binding to the CpG‐rich promoters of Pdcd4 and Pla2g6 . I,J) Western blot analysis of cleaved caspase‐3 levels in primary neurons expressing wild‐type MeCP2, or MeCP2 mutants under OGD/R conditions ( n = 4–7 per group). K–M) Western blot analysis of GVI PLA2 and PDCD4 protein levels in neurons expressing wild‐type MeCP2, or MeCP2 mutants under OGD/R conditions ( n = 7–8 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant.

Journal: Advanced Science

Article Title: MeCP2 Lactylation Protects against Ischemic Brain Injury by Transcriptionally Regulating Neuronal Apoptosis

doi: 10.1002/advs.202415309

Figure Lengend Snippet: Lactylation of MeCP2 at K210 and K249 regulates apoptotic gene expression in stroke. A) Sequence alignment of the transcriptional repression domain of MeCP2 across species, highlighting conserved MeCP2 lactylation sites. B) Co‐immunoprecipitation and Western blot analysis showing reduced MeCP2 lactylation levels upon K210R or K249R mutation. Luciferase reporter assays showing increased transcriptional activity at the C) Pdcd4 and D) Pla2g6 promoters in HEK293T cells expressing K210R or K249R MeCP2 mutants compared to wild‐type MeCP2 ( n = 3 per group). E) MeCP2 was immunoprecipitated with anti‐HA from HEK293T cells expressing WT or K210R/K249R mutants, and pan‐Kla and HA‐MeCP2 levels were detected by Western blot. Luciferase reporter assays showing transcriptional activity at the F) Pdcd4 and G) Pla2g6 promoters in HEK293T cells expressing WT MeCP2 or K210/K249 mutants ( n = 4 per group). H) EMSA analysis showing recombinant wild‐type and mutant MeCP2 binding to the CpG‐rich promoters of Pdcd4 and Pla2g6 . I,J) Western blot analysis of cleaved caspase‐3 levels in primary neurons expressing wild‐type MeCP2, or MeCP2 mutants under OGD/R conditions ( n = 4–7 per group). K–M) Western blot analysis of GVI PLA2 and PDCD4 protein levels in neurons expressing wild‐type MeCP2, or MeCP2 mutants under OGD/R conditions ( n = 7–8 per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant.

Techniques: Gene Expression, Sequencing, Immunoprecipitation, Western Blot, Mutagenesis, Luciferase, Activity Assay, Expressing, Recombinant, Binding Assay

Western-blot analysis (A) of calcium-dependent and independent phospholipase A2 (phosphor-cPLA2 and iPLA2, respectively), 4HNE levels and their quantification results (B, C, and D). (E) 4-HNE level measure by MDMS-shotgun lipidomics. Gene Ontology (GO) terms analysis of genes that negatively correlated with LPE in pharmacological (F) and genetic (G) microglial elimination cohort. Data transformation: square root for the pharmacological cohort, cube root for the genetic cohort; data scaling: pareto for the pharmacological cohort, mean for the genetic cohort. All data presented as mean ± SEM, normalized to WT. Two tailed two-way ANOVA with Turkey correction, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

Journal: bioRxiv

Article Title: Microglia-Dependent and Independent Modulation of Brain Lipid Metabolism in Alzheimer’s Disease Revealed by Pharmacological and Genetic Microglial Depletion

doi: 10.1101/2024.11.18.624173

Figure Lengend Snippet: Western-blot analysis (A) of calcium-dependent and independent phospholipase A2 (phosphor-cPLA2 and iPLA2, respectively), 4HNE levels and their quantification results (B, C, and D). (E) 4-HNE level measure by MDMS-shotgun lipidomics. Gene Ontology (GO) terms analysis of genes that negatively correlated with LPE in pharmacological (F) and genetic (G) microglial elimination cohort. Data transformation: square root for the pharmacological cohort, cube root for the genetic cohort; data scaling: pareto for the pharmacological cohort, mean for the genetic cohort. All data presented as mean ± SEM, normalized to WT. Two tailed two-way ANOVA with Turkey correction, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.

Article Snippet: PVDF membranes (CliniSciences, Nanterre, France) with the transferred protein were incubated with primary antibodies (1:1000-2000 dilutions) of anti-6E10 (mouse, BioLegend, Inc., San Diego, California, USA), anti-Homer1 (rabbit, Cell Signaling Technology, Boston, MA, USA), anti-CD68 (E307V) (rabbit, Cell Signaling Technology, Boston, MA, USA), anti-LAMP-1 (1D4B) (rat, Thermo Fisher Scientific, Waltham, MA, USA); anti-4HNE (12F7) (mouse, Thermo Fisher Scientific, Waltham, MA, USA), anti-PGRN (sheep, R&D Systems, Minneapolis, MN, USA), anti-Phospho-PLA2G4A (Ser505) (Rabbit, Proteintech Group, Inc, Rosemont, IL, USA), anti-iPLA2 (D-4) (mouse, Santa Cruz Biotechnology, Dallas, TX, USA), anti-HO-1 (rabbit, Proteintech Group, Inc, Rosemont, IL, USA), anti-Nrf2 (rabbit, Abcam, Waltham, MA, USA), anti-GFAP (rabbit, Thermo Fisher Scientific, Waltham, MA, USA), anti-GAPDH (rabbit, Cell Signaling Technology, USA) overnight at 4 °C, followed by horseradish peroxidase (HRP)-linked secondary antibodies (Cell Signaling Technology, Boston, MA, USA) for 1 h at room temperature.

Techniques: Western Blot, Transformation Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Microglia-Dependent and Independent Modulation of Brain Lipid Metabolism in Alzheimer’s Disease Revealed by Pharmacological and Genetic Microglial Depletion

doi: 10.1101/2024.11.18.624173

Techniques: Western Blot, Transformation Assay, Two Tailed Test

Overexpression of iPLA2β in PFC improves cognitive function of old mice. A Scheme of the experimental design. AAV injections were administered 21 days prior to formal behavioral testing. Behavioral test training sessions were conducted two days before the formal testing began. Samples were collected on the seventh day following the initiation of the formal behavioral experiment. B Protein levels of iPLA2β in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, as assessed by Western blotting and densitometry. C Representative immunofluorescence images of iPLA2β (red) and NeuN (green) in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Scale bar: 20 μm. D Representative SA-β-gal staining images in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Scale bar: 100 μm. E mRNA expression levels of TGFβ, IL-1β, TNF-α, and CCL2 in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups assessed via qPCR. Normalization was conducted relative to GAPDH expression levels. n = 11. F Representative IHC images of Iba1 in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Histogram showing quantification of the Iba1(+) area. Scale bar: 50 μm. G Travel time of the mice to reach the platform during the spatial test. n = 20 per group. H Duration spent by mice in the hidden platform quadrants during the probe trial. n = 20. I Frequency of platform crossings by mice during the probe trial. n = 20 per group. J Recognition index during the Novel object recognition test. Recognition index = time spent exploring a novel object/time spent exploring both objects. n = 20. K Discrimination index during the Novel object recognition test. The discrimination index = (time spent on the novel object − time spent on the familiar object)/ time spent on both objects. n = 20 Data are presented as the mean ± SEM; p values were obtained using the Mann-Whitney U test (B, C, D, and F) and the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (E, G, H, I, J, and K). * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: Overexpression of iPLA2β in PFC improves cognitive function of old mice. A Scheme of the experimental design. AAV injections were administered 21 days prior to formal behavioral testing. Behavioral test training sessions were conducted two days before the formal testing began. Samples were collected on the seventh day following the initiation of the formal behavioral experiment. B Protein levels of iPLA2β in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, as assessed by Western blotting and densitometry. C Representative immunofluorescence images of iPLA2β (red) and NeuN (green) in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Scale bar: 20 μm. D Representative SA-β-gal staining images in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Scale bar: 100 μm. E mRNA expression levels of TGFβ, IL-1β, TNF-α, and CCL2 in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups assessed via qPCR. Normalization was conducted relative to GAPDH expression levels. n = 11. F Representative IHC images of Iba1 in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups. Histogram showing quantification of the Iba1(+) area. Scale bar: 50 μm. G Travel time of the mice to reach the platform during the spatial test. n = 20 per group. H Duration spent by mice in the hidden platform quadrants during the probe trial. n = 20. I Frequency of platform crossings by mice during the probe trial. n = 20 per group. J Recognition index during the Novel object recognition test. Recognition index = time spent exploring a novel object/time spent exploring both objects. n = 20. K Discrimination index during the Novel object recognition test. The discrimination index = (time spent on the novel object − time spent on the familiar object)/ time spent on both objects. n = 20 Data are presented as the mean ± SEM; p values were obtained using the Mann-Whitney U test (B, C, D, and F) and the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (E, G, H, I, J, and K). * p < 0.05. **, p < 0.01; ***, p < 0.001

Article Snippet: The primary antibodies employed in this study included iPLA2β (sc-376563, 1:100, Santa Cruz), NeuN (ab177487, 1:200, abcam), Iba1 (#17198, 1:400, Cell Signaling), GFAP (ab7260, 1:500, Abcam) and TOM20 (A19403, 1:200, ABclonal).

Techniques: Over Expression, Control, Western Blot, Immunofluorescence, Staining, Expressing, MANN-WHITNEY

Aging increased iPLA2β loss in the PFC of mice. A PLA2s mRNA expression levels in the PFC were assessed using qPCR. Normalization was conducted relative to GAPDH expression levels. n = 10. B iPLA2β protein levels in the PFC were assessed via Western blotting. n = 3. C Representative IHC images depicting iPLA2β in the PFC. IHC analysis showing the density of iPLA2β (+) cells/mm². Scale bar: 100 μm. n = 16. D Representative immunofluorescence images depicting iPLA2β (red) and NeuN (green) in the PFC. The bar graph shows the percentage of iPLA2β(+) NeuN(+) cells relative to the total NeuN(+) cell population (%).Scale bar: 20 μm. n = 10 M represents “month” Data are presented as mean ± SEM; p values were obtained using Mann-Whitney U test (A), one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (B), and two-sided unpaired Student’s t-tests (C, D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: Aging increased iPLA2β loss in the PFC of mice. A PLA2s mRNA expression levels in the PFC were assessed using qPCR. Normalization was conducted relative to GAPDH expression levels. n = 10. B iPLA2β protein levels in the PFC were assessed via Western blotting. n = 3. C Representative IHC images depicting iPLA2β in the PFC. IHC analysis showing the density of iPLA2β (+) cells/mm². Scale bar: 100 μm. n = 16. D Representative immunofluorescence images depicting iPLA2β (red) and NeuN (green) in the PFC. The bar graph shows the percentage of iPLA2β(+) NeuN(+) cells relative to the total NeuN(+) cell population (%).Scale bar: 20 μm. n = 10 M represents “month” Data are presented as mean ± SEM; p values were obtained using Mann-Whitney U test (A), one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (B), and two-sided unpaired Student’s t-tests (C, D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques: Expressing, Western Blot, Immunofluorescence, MANN-WHITNEY

Deficiency of iPLA2β increases aging-related cellular senescence, cognitive impairment and neuroinflammation. A The mRNA expression levels of iPLA2β in the PFC of 24 M mice were assessed by qPCR. Normalization was performed relative to GAPDH expression levels. n = 6. B Protein levels of iPLA2β in the PFC of 24 M mice, assessed using Western blotting and densitometry, n = 3. C Time taken by 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice to reach the platform during the spatial test. n = 20. D Frequency of platform crossings in 24 M mice during the probe trial. n = 20. E Duration of 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice spent time in hidden platform quadrants during the probe trial. n = 20. F Recognition index of 2 M-WT, 24 M-WT, 24 M-CON and 24 M-KO mice during the Novel object recognition test. Recognition index = time spent exploring novel object/time spent exploring both objects. n = 20. G The discrimination index of 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice during the Novel object recognition test. The discrimination index = (time spent on the novel object − time spent on the familiar object)/ time spent on both objects. n = 20. H Representative SA-β-gal staining images in 24 M PFC. The bar graph shows the density of β-Gal staining (+) cells/0.1 mm². Scale bar: 100 μm. n = 10. I mRNA levels of TGF-β, CCL2, TNF-α, and IL-1β in the PFC of 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice were assessed via qPCR. Normalization was conducted relative to GAPDH expression levels. n = 11 KO represents “iPLA2β knockout,” CON represents “control”, Data are presented as mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B, H), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (C, D, E, F, G and I), * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: Deficiency of iPLA2β increases aging-related cellular senescence, cognitive impairment and neuroinflammation. A The mRNA expression levels of iPLA2β in the PFC of 24 M mice were assessed by qPCR. Normalization was performed relative to GAPDH expression levels. n = 6. B Protein levels of iPLA2β in the PFC of 24 M mice, assessed using Western blotting and densitometry, n = 3. C Time taken by 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice to reach the platform during the spatial test. n = 20. D Frequency of platform crossings in 24 M mice during the probe trial. n = 20. E Duration of 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice spent time in hidden platform quadrants during the probe trial. n = 20. F Recognition index of 2 M-WT, 24 M-WT, 24 M-CON and 24 M-KO mice during the Novel object recognition test. Recognition index = time spent exploring novel object/time spent exploring both objects. n = 20. G The discrimination index of 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice during the Novel object recognition test. The discrimination index = (time spent on the novel object − time spent on the familiar object)/ time spent on both objects. n = 20. H Representative SA-β-gal staining images in 24 M PFC. The bar graph shows the density of β-Gal staining (+) cells/0.1 mm². Scale bar: 100 μm. n = 10. I mRNA levels of TGF-β, CCL2, TNF-α, and IL-1β in the PFC of 2 M-WT, 24 M-WT, 24 M-CON, and 24 M-KO mice were assessed via qPCR. Normalization was conducted relative to GAPDH expression levels. n = 11 KO represents “iPLA2β knockout,” CON represents “control”, Data are presented as mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B, H), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (C, D, E, F, G and I), * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques: Expressing, Western Blot, Staining, Knock-Out, Control

iPLA2β reduces senescence in primary neurons. A The mRNA levels of iPLA2β in DIV7 and DIV20 cultured neurons were assessed using qPCR. Normalization was conducted relative to GAPDH expression levels. n = 13. B Protein levels of iPLA2β in DIV7 and DIV20 cultured primary neurons, assessed via Western blot and densitometry. n = 4. C Representative SA-β-gal staining images of iPLA2β-overexpression (OE) and control DIV20 primary neurons. Scale bar: 100 μm. n = 12. D Representative immunofluorescence images of iPLA2β (red) in D-gal-induced iPLA2β overexpression (OE) and control primary neurons. Scale bar: 5 μm. n = 8. E Representative SA-β-gal staining images of D-gal-induced iPLA2β-overexpression (OE) and control primary neurons. Scale bar: 100 μm. n = 12. F Protein levels of P62 and P16 in D-gal-induced iPLA2β-overexpression (OE) and control primary neurons, as assessed by Western blot and densitometry OE represents “iPLA2β overexpression”, CON represents “control”, Data are mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B), Mann-Whitney U test (C), one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (E), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (F), * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: iPLA2β reduces senescence in primary neurons. A The mRNA levels of iPLA2β in DIV7 and DIV20 cultured neurons were assessed using qPCR. Normalization was conducted relative to GAPDH expression levels. n = 13. B Protein levels of iPLA2β in DIV7 and DIV20 cultured primary neurons, assessed via Western blot and densitometry. n = 4. C Representative SA-β-gal staining images of iPLA2β-overexpression (OE) and control DIV20 primary neurons. Scale bar: 100 μm. n = 12. D Representative immunofluorescence images of iPLA2β (red) in D-gal-induced iPLA2β overexpression (OE) and control primary neurons. Scale bar: 5 μm. n = 8. E Representative SA-β-gal staining images of D-gal-induced iPLA2β-overexpression (OE) and control primary neurons. Scale bar: 100 μm. n = 12. F Protein levels of P62 and P16 in D-gal-induced iPLA2β-overexpression (OE) and control primary neurons, as assessed by Western blot and densitometry OE represents “iPLA2β overexpression”, CON represents “control”, Data are mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B), Mann-Whitney U test (C), one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (E), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (F), * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques: Cell Culture, Expressing, Western Blot, Staining, Over Expression, Control, Immunofluorescence, MANN-WHITNEY

iPLA2β protects mitochondrial morphology and regulate mitochondrial function during cortex aging. A Representative TEM image of WT and iPLA2β −/− PFC of 24 M mice. Scale bar: 1 μm. B Representative TEM images of the AAV-CON-injected and AAV-iPLA2β-OE-injected PFC of mice. Scale bar: 1 μm. C Quantification of mitochondrial diameter in TEM images of WT and iPLA2β −/− PFC from 24 M mice. n = 10. D Quantification of mitochondrial diameter in TEM images of the AAV-CON-injected and AAV- iPLA2β-OE-injected PFC of mice. n = 10. E Quantification of mitochondrial length in TEM images of WT and iPLA2β −/− PFC from 24 M mice. n = 10. F Quantification of mitochondrial length in TEM images of the AAV-CON-injected and AAV- iPLA2β-OE-injected PFC of mice. n = 10. G ATP levels in the WT and iPLA2β −/− PFC of 24 M mice. n = 8. H ATP levels of the AAV-CON-injected and AAV- iPLA2β-OE-injected PFC of mice. n = 8. I Representative immunofluorescence images of iPLA2β (green) and TOM20 (red) in the PFC of 2 M-WT, 24 M-WT, AAV-CON-injected, and AAV- iPLA2β-OE-injected mice. Scale bar: 50 μm. J TOM20 fluorescence intensity of immunofluorescence images in (I). n = 12. K iPLA2β fluorescence intensity of immunofluorescence images in (I). n = 12 KO represents “iPLA2β knockout,” WT represents “wildtype”, OE represents “iPLA2β overexpression”, CON represents “control”, Data are mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (C, D, E, F, G, and H), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (J and K), * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: iPLA2β protects mitochondrial morphology and regulate mitochondrial function during cortex aging. A Representative TEM image of WT and iPLA2β −/− PFC of 24 M mice. Scale bar: 1 μm. B Representative TEM images of the AAV-CON-injected and AAV-iPLA2β-OE-injected PFC of mice. Scale bar: 1 μm. C Quantification of mitochondrial diameter in TEM images of WT and iPLA2β −/− PFC from 24 M mice. n = 10. D Quantification of mitochondrial diameter in TEM images of the AAV-CON-injected and AAV- iPLA2β-OE-injected PFC of mice. n = 10. E Quantification of mitochondrial length in TEM images of WT and iPLA2β −/− PFC from 24 M mice. n = 10. F Quantification of mitochondrial length in TEM images of the AAV-CON-injected and AAV- iPLA2β-OE-injected PFC of mice. n = 10. G ATP levels in the WT and iPLA2β −/− PFC of 24 M mice. n = 8. H ATP levels of the AAV-CON-injected and AAV- iPLA2β-OE-injected PFC of mice. n = 8. I Representative immunofluorescence images of iPLA2β (green) and TOM20 (red) in the PFC of 2 M-WT, 24 M-WT, AAV-CON-injected, and AAV- iPLA2β-OE-injected mice. Scale bar: 50 μm. J TOM20 fluorescence intensity of immunofluorescence images in (I). n = 12. K iPLA2β fluorescence intensity of immunofluorescence images in (I). n = 12 KO represents “iPLA2β knockout,” WT represents “wildtype”, OE represents “iPLA2β overexpression”, CON represents “control”, Data are mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (C, D, E, F, G, and H), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (J and K), * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques: Injection, Immunofluorescence, Fluorescence, Knock-Out, Over Expression, Control

iPLA2β regulates mitophagy during neuronal aging in vivo. A Quantification of the mtDNA/nDNA ratio in the 24 M PFC in the wild-type (WT) and iPLA2β knockout (iPLA2β −/− ) groups, using 16 S rRNA and Hexokinase 2 (Hk2), respectively. n = 6. B Optineurin, PINK1, Parkin, MFF, and LC3B levels in the 24 M PFC in wild-type (WT) and iPLA2β knockout (iPLA2β −/− ) groups, assessed by Western blot and densitometry. n = 3. C BNIP3 and NIX levels in the 24 M PFC in wild-type (WT) and iPLA2β knockout (iPLA2β −/− ) groups, assessed by Western blot and densitometry. n = 3. D Quantification of the mtDNA/nDNA ratio in the 22 M PFC with iPLA2β overexpression (OE) and control (CON) groups, using 16 S rRNA and Hexokinase 2 (Hk2), respectively. n = 6. E Optineurin, PINK1, Parkin, MFF, and LC3B levels in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, assessed by Western blot and densitometry. n = 3. F BNIP3 and NIX levels in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, as assessed by Western blotting and densitometry. n = 3 KO represents “iPLA2β knockout”, WT represents “wildtype”, OE represents “iPLA2β overexpression”, CON represents “control”, Data are presented as the mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B, C, and D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: iPLA2β regulates mitophagy during neuronal aging in vivo. A Quantification of the mtDNA/nDNA ratio in the 24 M PFC in the wild-type (WT) and iPLA2β knockout (iPLA2β −/− ) groups, using 16 S rRNA and Hexokinase 2 (Hk2), respectively. n = 6. B Optineurin, PINK1, Parkin, MFF, and LC3B levels in the 24 M PFC in wild-type (WT) and iPLA2β knockout (iPLA2β −/− ) groups, assessed by Western blot and densitometry. n = 3. C BNIP3 and NIX levels in the 24 M PFC in wild-type (WT) and iPLA2β knockout (iPLA2β −/− ) groups, assessed by Western blot and densitometry. n = 3. D Quantification of the mtDNA/nDNA ratio in the 22 M PFC with iPLA2β overexpression (OE) and control (CON) groups, using 16 S rRNA and Hexokinase 2 (Hk2), respectively. n = 6. E Optineurin, PINK1, Parkin, MFF, and LC3B levels in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, assessed by Western blot and densitometry. n = 3. F BNIP3 and NIX levels in the 22 M PFC with iPLA2β overexpression (iPLA2β-OE) and control (CON) groups, as assessed by Western blotting and densitometry. n = 3 KO represents “iPLA2β knockout”, WT represents “wildtype”, OE represents “iPLA2β overexpression”, CON represents “control”, Data are presented as the mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B, C, and D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques: In Vivo, Knock-Out, Western Blot, Over Expression, Control

iPLA2β regulates mitophagy during neuronal aging in vitro. A Primary cultured cortical neurons were transfected with GFP-LC3B and Mito-DsRed. Fluorescent images were captured 3 h after reperfusion. Scale bar: 5 μm. B Relative colocalization ratio between GFP-LC3B and Mito-DsRed in immunofluorescence images in (A). The ratio was calculated by dividing the number of LC3B-Mito puncta by the total number of Mito puncta. n = 6. C Mitochondrial levels of iPLA2β, Parkin, optineurin, and PINK1 were assessed by Western blot and densitometry in D-gal-induced iPLA2β overexpression (OE) and control primary neurons. D Protein levels of MFF and LC3B were evaluated using Western blot and densitometry in D-gal-induced iPLA2β overexpression (OE) and control primary neurons CQ represents “chloroquine”, D-gal represents “D-galactose”, OE represents “iPLA2β overexpression”, NC represents “Negative Control”, Data are presented as the mean ± SEM; p values were obtained using, one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (B), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (C and D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: iPLA2β regulates mitophagy during neuronal aging in vitro. A Primary cultured cortical neurons were transfected with GFP-LC3B and Mito-DsRed. Fluorescent images were captured 3 h after reperfusion. Scale bar: 5 μm. B Relative colocalization ratio between GFP-LC3B and Mito-DsRed in immunofluorescence images in (A). The ratio was calculated by dividing the number of LC3B-Mito puncta by the total number of Mito puncta. n = 6. C Mitochondrial levels of iPLA2β, Parkin, optineurin, and PINK1 were assessed by Western blot and densitometry in D-gal-induced iPLA2β overexpression (OE) and control primary neurons. D Protein levels of MFF and LC3B were evaluated using Western blot and densitometry in D-gal-induced iPLA2β overexpression (OE) and control primary neurons CQ represents “chloroquine”, D-gal represents “D-galactose”, OE represents “iPLA2β overexpression”, NC represents “Negative Control”, Data are presented as the mean ± SEM; p values were obtained using, one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (B), the Kruskal-Wallis test followed by Dunn’s multiple comparisons test (C and D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques: In Vitro, Cell Culture, Transfection, Immunofluorescence, Western Blot, Over Expression, Control, Negative Control

iPLA2β deficiency leads to alterations in mitochondrial phospholipid metabolism in aged PFC. A Heat map representing individual LPC species and lipidomic analysis of LPC species were significantly altered in the KO group in 24 M PFCs. B Heat map representing individual LPE species and lipidomic analysis of LPE species significantly altered in the KO group in 24 M PFCs. C Heat map representing individual PC species and lipidomic analysis of PC species significantly altered in the KO group in 24 M PFCs. D Heat map representing individual PE species and lipidomic analysis of PE species significantly altered in the KO group in 24 M PFCs MUFA represents “monounsaturated fatty acid”; PUFA represents “polyunsaturated fatty acid”. Data are presented as the mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B, C, and D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: iPLA2β deficiency leads to alterations in mitochondrial phospholipid metabolism in aged PFC. A Heat map representing individual LPC species and lipidomic analysis of LPC species were significantly altered in the KO group in 24 M PFCs. B Heat map representing individual LPE species and lipidomic analysis of LPE species significantly altered in the KO group in 24 M PFCs. C Heat map representing individual PC species and lipidomic analysis of PC species significantly altered in the KO group in 24 M PFCs. D Heat map representing individual PE species and lipidomic analysis of PE species significantly altered in the KO group in 24 M PFCs MUFA represents “monounsaturated fatty acid”; PUFA represents “polyunsaturated fatty acid”. Data are presented as the mean ± SEM; p values were obtained using two-sided unpaired Student’s t-tests (A, B, C, and D). * p < 0.05. **, p < 0.01; ***, p < 0.001

Techniques:

Possible mechanism underlying the protective effects of iPLA2β on brain aging

Journal: Journal of Neuroinflammation

Article Title: iPLA2β loss leads to age-related cognitive decline and neuroinflammation by disrupting neuronal mitophagy

doi: 10.1186/s12974-024-03219-z

Figure Lengend Snippet: Possible mechanism underlying the protective effects of iPLA2β on brain aging

Techniques:

Review

Structured Review

Images

Similar Products

Image Search Results