Journal: The FASEB Journal

Article Title: Polyunsaturated fatty acid deficiency affects sulfatides and other sulfated glycans in lysosomes through autophagy‐mediated degradation

doi: 10.1096/fj.202000030rr

Figure Lengend Snippet: FIGURE 1 Changes in sulfatide levels and the expression of sulfatide metabolic enzymes in the brain of mice. A, Sulfatide levels in the brain were measured by MALDI-TOF MS. The total amount of sulfatides was calculated as the sum of seven lysosulfatide molecular species (μmol/g wet brain weight). PUFA (+), group supplemented with PUFA-containing oil; PUFA (−), group supplemented with PUFA-deficient oil. Data are expressed as the mean ± SD (n = 3-4). P values comparing each time point with control mice of 0 weeks were calculated by the unpaired Student's t test: §§§P < .001. Differences between groups were compared using ANOVA with Tukey's post hoc test: **P < .01, ***P < .001 vs PUFA (+) group; ##P < .01, ###P < .001 vs control group. B, Immunoblot and gray analyses showing the expressions of protein levels of sulfatide- synthesizing enzymes (CST and CGT) and sulfatide-degrading enzymes (ARSA and GALC). 5w HCO: mice fed a 14% HCO-based diet for 5 weeks. Protein levels were normalized to that of β-actin. Analysis of mRNA levels of these enzymes were performed by real-time PCR and normalized to that of GAPDH. All data are shown as fold changes relative to the control group. Data are expressed as the mean ± SD (n = 3-4). Statistically significant differences: **P < .01, ***P < .001 vs PUFA (+) group; ##P < .01, ###P < .001 vs control group

Article Snippet: The preparation of whole-tissue lysates (brain and kidney) and immunoblot analysis were conducted as described previously.32,33 The primary antibodies used for immunoblot testing included anti-β-actin (1:1000), anti-cerebroside sulfotransferase (CST) (1:2000), anti-ceramide galactosyltransferase (CGT) (1:2000), anti-arylsulfatase A (ARSA) (1:2000), anti-mammalian target of rapamycin (mTOR) (1:1000), anti-phospho-mTOR (S2448) (1:1000), anti-AKT (1:4000), and anti-amyloid precursor protein (APP) (1:5000) | 9597WANG et Al. (Abcam, Cambridge, UK); anti-galactosylceramidase (GALC) (1:1000) (Proteintech, Chicago, IL, USA); antip62 (1:1000), anti-Beclin1 (1:1000), and anti-Atg5 (1:500) (Medical & Biological Laboratories, Co., Ltd., Nagoya, Japan); anti-microtubule associated protein 1 light chain 3B (LC3B) (1:500) (Novus Biologicals, Centennial, CO, USA); anti-P53 (1:500) (Santa Cruz Biotechnology, Dallas, TX, USA); anti-extracellular regulated protein kinase-1/2 (Erk1/2) (1:1000) (Upstate Cell Signaling Solution, Lake Placid, NY, USA); anti-phospho-Erk1/2 (1:2000), anti-AMP-activated protein kinase α (AMPKα) (1:1000), anti-phospho-AMPKα (1:1000), and anti-phospho-AKT (1:1000) (Cell Signaling Technology, Danvers, MA, USA); and anti-β-glucuronidase (1:500)34 that was kindly provided by Dr K. Sukegawa from Gifu University School of Medicine, Gifu, Japan.

Techniques: Expressing, Control, Western Blot, Real-time Polymerase Chain Reaction

Journal: The FASEB Journal

Article Title: Polyunsaturated fatty acid deficiency affects sulfatides and other sulfated glycans in lysosomes through autophagy‐mediated degradation

doi: 10.1096/fj.202000030rr

Figure Lengend Snippet: FIGURE 3 Time-course changes in expression levels of ARSA and LC3B in the brain of mice. The protein expression levels of ARSA and LC3B were examined by immunoblot and gray analyses. Protein levels were normalized to that of β-actin. All data are shown as fold changes relative to the control group at 0 weeks. Data are expressed as the mean ± SD (n = 3-4). P values comparing each time point with control mice of 0 weeks were calculated by the unpaired Student's t test: §P < .05, §§P < .01, §§§P < .001

Article Snippet: The preparation of whole-tissue lysates (brain and kidney) and immunoblot analysis were conducted as described previously.32,33 The primary antibodies used for immunoblot testing included anti-β-actin (1:1000), anti-cerebroside sulfotransferase (CST) (1:2000), anti-ceramide galactosyltransferase (CGT) (1:2000), anti-arylsulfatase A (ARSA) (1:2000), anti-mammalian target of rapamycin (mTOR) (1:1000), anti-phospho-mTOR (S2448) (1:1000), anti-AKT (1:4000), and anti-amyloid precursor protein (APP) (1:5000) | 9597WANG et Al. (Abcam, Cambridge, UK); anti-galactosylceramidase (GALC) (1:1000) (Proteintech, Chicago, IL, USA); antip62 (1:1000), anti-Beclin1 (1:1000), and anti-Atg5 (1:500) (Medical & Biological Laboratories, Co., Ltd., Nagoya, Japan); anti-microtubule associated protein 1 light chain 3B (LC3B) (1:500) (Novus Biologicals, Centennial, CO, USA); anti-P53 (1:500) (Santa Cruz Biotechnology, Dallas, TX, USA); anti-extracellular regulated protein kinase-1/2 (Erk1/2) (1:1000) (Upstate Cell Signaling Solution, Lake Placid, NY, USA); anti-phospho-Erk1/2 (1:2000), anti-AMP-activated protein kinase α (AMPKα) (1:1000), anti-phospho-AMPKα (1:1000), and anti-phospho-AKT (1:1000) (Cell Signaling Technology, Danvers, MA, USA); and anti-β-glucuronidase (1:500)34 that was kindly provided by Dr K. Sukegawa from Gifu University School of Medicine, Gifu, Japan.

Techniques: Expressing, Western Blot, Control

Journal: The FASEB Journal

Article Title: Polyunsaturated fatty acid deficiency affects sulfatides and other sulfated glycans in lysosomes through autophagy‐mediated degradation

doi: 10.1096/fj.202000030rr

Figure Lengend Snippet: FIGURE 8 Time-course changes in the expression levels of ARSA and LC3B in the kidney. The protein expression levels of ARSA and LC3B were examined by immunoblot and gray analyses. Protein levels were normalized to that of β-actin. All data are shown as fold changes relative to the control group at 0 weeks. Data are expressed as the mean ± SD (n = 3-4). P values comparing each time point with control mice of 0 weeks were calculated by the unpaired Student's t test: §P < .05

Article Snippet: The preparation of whole-tissue lysates (brain and kidney) and immunoblot analysis were conducted as described previously.32,33 The primary antibodies used for immunoblot testing included anti-β-actin (1:1000), anti-cerebroside sulfotransferase (CST) (1:2000), anti-ceramide galactosyltransferase (CGT) (1:2000), anti-arylsulfatase A (ARSA) (1:2000), anti-mammalian target of rapamycin (mTOR) (1:1000), anti-phospho-mTOR (S2448) (1:1000), anti-AKT (1:4000), and anti-amyloid precursor protein (APP) (1:5000) | 9597WANG et Al. (Abcam, Cambridge, UK); anti-galactosylceramidase (GALC) (1:1000) (Proteintech, Chicago, IL, USA); antip62 (1:1000), anti-Beclin1 (1:1000), and anti-Atg5 (1:500) (Medical & Biological Laboratories, Co., Ltd., Nagoya, Japan); anti-microtubule associated protein 1 light chain 3B (LC3B) (1:500) (Novus Biologicals, Centennial, CO, USA); anti-P53 (1:500) (Santa Cruz Biotechnology, Dallas, TX, USA); anti-extracellular regulated protein kinase-1/2 (Erk1/2) (1:1000) (Upstate Cell Signaling Solution, Lake Placid, NY, USA); anti-phospho-Erk1/2 (1:2000), anti-AMP-activated protein kinase α (AMPKα) (1:1000), anti-phospho-AMPKα (1:1000), and anti-phospho-AKT (1:1000) (Cell Signaling Technology, Danvers, MA, USA); and anti-β-glucuronidase (1:500)34 that was kindly provided by Dr K. Sukegawa from Gifu University School of Medicine, Gifu, Japan.

Techniques: Expressing, Western Blot, Control

Journal: Molecular genetics & genomic medicine

Article Title: Identification of a missense ARSA mutation in metachromatic leukodystrophy and its potential pathogenic mechanism.

doi: 10.1002/mgg3.1478

Figure Lengend Snippet: FIGURE 2 Multiple sequence alignment (MSA) and 3D structure of ARSA. (a) Multiple sequence alignment showing the sequence alignment of a specific amino acid, and its conservation in other ARSA orthologs (across different species). Nucleotide numbers are derived from cDNA ARSA sequences, GenBank accession numbers: NM_000487.5 and NP_000478.3. (b) Conformational changes induced by the p.E309K and p.E309Q missense mutation in the ARSA protein. (c) Conformational changes induced by the p.E309*, this mutation results in the early termination of codons and truncated proteins.

Article Snippet: 2.3 | Overexpression cell models of the mutated ARSA gene The human wild-type ARSA cDNA (cloned in the pCMV6 plasmid) was purchased from OriGene (Cat. No. RC204319, OriGene, USA).

Techniques: Sequencing, Derivative Assay, Mutagenesis

Journal: Molecular genetics & genomic medicine

Article Title: Identification of a missense ARSA mutation in metachromatic leukodystrophy and its potential pathogenic mechanism.

doi: 10.1002/mgg3.1478

Figure Lengend Snippet: FIGURE 1 Proband, family, and mutation. (A) MRI from the proband (II-2). Magnetic resonance imaging (MRI) shows symmetrical deep lesions located in periventricular white matter, which was low signal in T1WI (a), high signal in T2WI (b), low signal in in T2WI (c) and ep2d (d) from the proband (II-2). (B) Pedigree of the family with MLD patients. The proband was shown in the second generation with the numbers II-2. The parents of proband are first generation with the number I-1 and I-2. The healthy older brother of proband is in the second generation with the number II-1. (C) Mutational analysis of the arylsulfatase A (ARSA) gene. Genotypes of the proband showed a homozygous c.925G>A mutation, and those of the parents showed a heterozygous c.925G>A mutation. His healthy brother did not inherit this mutation. Nucleotide numbers are derived from cDNA ARSA sequences, GenBank accession numbers: NM_000487.5 and NP_000478.3.

Article Snippet: 2.3 | Overexpression cell models of the mutated ARSA gene The human wild-type ARSA cDNA (cloned in the pCMV6 plasmid) was purchased from OriGene (Cat. No. RC204319, OriGene, USA).

Techniques: Mutagenesis, Magnetic Resonance Imaging, Derivative Assay

Journal: Molecular genetics & genomic medicine

Article Title: Identification of a missense ARSA mutation in metachromatic leukodystrophy and its potential pathogenic mechanism.

doi: 10.1002/mgg3.1478

Figure Lengend Snippet: FIGURE 3 Transcriptomic analysis in overexpression cell models of wild-type and mutated ARSA gene. (a) Construction of overexpression cell models of wild-type and mutated ARSA gene. (b) Relative mRNA expression of ARSA gene, GAPDH was used as a loading control. *p < 0.05 in independent Student's t-test, ***p < 0.001 in independent Student's t-test (n = 3). (c) Sulfatides concentration in wild-type cells and ARSA gene overexpression cell models. The standardization of sulfatides amount was performed by dividing it into the amount of total protein of the lysates. No significant difference was found in independent Student's t-test (n = 3). (d) Hot Map of Characteristic Gene Adjacency in Characteristic Gene Network. Each row and column corresponds to a characteristic gene (marked with the same color). In heatmap, red denotes high adjacency (positive correlation) and blue denotes low adjacency (negative correlation), as shown in the color legend. (e) Characteristic gene module-trait association of each module. Each row corresponds to a module characteristic gene, and each column corresponds to a cell type (trait). Each cell contains Pearson correlation coefficients (numbers outside parentheses) and associated p values (numbers inside parentheses). According to the color legend, color coding is carried out by correlation. Red indicates positive correlation and blue indicates a negative correlation.

Article Snippet: 2.3 | Overexpression cell models of the mutated ARSA gene The human wild-type ARSA cDNA (cloned in the pCMV6 plasmid) was purchased from OriGene (Cat. No. RC204319, OriGene, USA).

Techniques: Over Expression, Expressing, Control, Concentration Assay

Journal: Molecular genetics & genomic medicine

Article Title: Identification of a missense ARSA mutation in metachromatic leukodystrophy and its potential pathogenic mechanism.

doi: 10.1002/mgg3.1478

Figure Lengend Snippet: FIGURE 4 Protein–protein interaction networks in the overexpression cell models of wild-type and mutated ARSA gene.

Article Snippet: 2.3 | Overexpression cell models of the mutated ARSA gene The human wild-type ARSA cDNA (cloned in the pCMV6 plasmid) was purchased from OriGene (Cat. No. RC204319, OriGene, USA).

Techniques: Over Expression

Journal: Acta Pharmaceutica Sinica. B

Article Title: Next-generation aluminum adjuvants: Immunomodulatory layered double hydroxide NanoAlum reengineered from first-line drugs

doi: 10.1016/j.apsb.2024.09.012

Figure Lengend Snippet: LDH NanoAlum promote antitumor therapeutic effects in response to external stimuli. (A) therapeutic effects induced by FeOOH@STA/Cu-LDH nanohybrid through virtue of a unique ICD maximization strategy, to maximize primary 4T1 tumoral ICD along with magnificent CRT expression, thereafter awakening CTL for systemic tumor immune elimination. Reprinted with permission from Ref. . Copyright © 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. (B) The synthetic procedure of MLD NPs and their self-enhanced SDT and CDT effect. Reprinted with permission from Ref. . Copyright © 2022 Wiley-VCH GmbH. (C) The construction of Ca 2+ -introduced MgCaFe-LDH to fulfil high-performance oxidation stress and outstanding anti-tumor immunotherapy by activating T-cell mediated immunity. Reprinted with permission from Ref. . Copyright © 2023 Elsevier Ltd. (D) The preparation of a-Mn-CoMo-LDH-PEG as a sonosensitizer for MRI-guided sonodynamic cancer therapy. Reprinted with permission from Ref. 115 Copyright © 2023 Elsevier B.V.

Article Snippet: Copyright © 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. (B) The synthetic procedure of MLD NPs and their self-enhanced SDT and CDT effect.

Techniques: Expressing