Journal: iScience

Article Title: Targeting HOTAIRM1 ameliorates glioblastoma by disrupting mitochondrial oxidative phosphorylation and serine metabolism

doi: 10.1016/j.isci.2022.104823

Figure Lengend Snippet: Screening and identification of the target genes of HOTAIRM1 (A) Schematic outline of the process used to screen for HOTAIRM1 target genes in various GBM cell lines. (B) Chart showing qPCR verification of the 32 candidate target genes. Compared with shNT, data are presented as means and SDs. (C and D) Western blot analysis showing the changes in protein expression for the 15 candidate target genes. β-Actin was used as the loading control. (E) Western blot analysis showing SHMT1 protein levels. (F) qPCR analysis of SHMT2 in human astrocytes and glioma cell lines. Data are presented as means ± SDs. (G) Spearman correlation analysis of HOTAIRM1 and SHMT2 mRNA in the brain cortex and GBM samples with the GEPIA web server. (H) Statistical analysis of SHMT2 protein levels in glioma tissues compared with normal brain tissues. Data are presented as means ± SDs (∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA with unpaired t test). (I) Correlations analysis of the two HOTAIRM1 variants and the SHMT2 protein in normal brain and glioma tissues (n = 34). (J and K) SHMT2 knockdown xenograft tumors (shSHMT2) compared to control tumors (shNT) are pictured (J), tumor weights and volumes (K) were shown. n = 5 in each group]. Data are presented as means ± SDs (Compared with shNT, ∗p < 0.05, 2-tailed Student’s t test). (L–N) After knockdown of SHMT2 (L), the glycine/L-serine (M) and THF/5,10-CH 2 -THF (N) concentrations in A172 cells cultured with complete medium or serine/glycine-deprived medium were measured by UHPLC-MRM-MS. Data (M and N) are presented as means ± SDs (Compared with shNT, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA with Dunnett t test). N.D.: the target compound was not detected. See also Figures S2 , , Tables S1 and .

Article Snippet: The other human astrocyte lines HA (from the cerebral cortex), HAc (from the cerebellum), and HAsp (from the spinal cord) were purchased from ScienCell Research Laboratories and cultured with astrocyte medium (catalog no. 1801).

Techniques: Western Blot, Expressing, Control, Knockdown, Cell Culture

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: Validation of PA treatment and effects of PA treatment on oxidative stress markers in the HBAs. ( A , B ): changes in mRNA levels of lipid droplet associated genes, namely, Plin1 and Plin2 in the human brainstem astrocytes between control and PA-treated group. ( C ): Oil Red-O staining of the control and PA-treated cells at 20X. Scale bar indicates 100 μm. Data are means ± SEM ( n = 4–6 replicates per group). * significant difference ( p < 0.05) between the groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Biomarker Discovery, Control, Staining

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: Effects of PA treatment on oxidative stress markers in HBAs. ( A – C ): changes in mRNA levels of oxidative stress markers NQO1, SOD2, and CAT in human brainstem between control and PA-treated groups. ( D ): dihydroethidium staining of the control and PA-treated group with bar graphs for fluorescence intensity. Data are means ± SEM ( n = 3–6 replicates per group). Scale bar indicates 100 μm. * significant difference ( p < 0.05) between groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Control, Staining, Fluorescence

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: Immunofluorescence staining of Gamma H2AX (γ-H2AX) in the HBAs. ( A ): immunofluorescence staining for the DNA damage marker γ-H2AX in the human brainstem astrocytes between control and PA-treated groups. ( B ): bar graphs showing fluorescence intensity of γ-H2AX between the control and PA-treated groups. Data are means ± SEM ( n = 4 replicates per group). Scale bar indicates 100 μm. * significant difference ( p < 0.05) between groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Immunofluorescence, Staining, Marker, Control, Fluorescence

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: PA-induced oxidative stress causes cellular senescence in the HBAs. ( A – D ): Changes in the mRNA levels of senescence markers p16, p21, p53 and the nuclear envelope gene Lamin B1 in the human brainstem astrocytes between control and PA-treated groups. Data are means ± SEM ( n = 5–6 replicates per group). * Significant difference ( p < 0.05) between the groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Control

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: Immunofluorescence staining of p16 INK4a in the HBAs. ( A ): immunofluorescence staining for the senescence marker p16 INK4a in the human brainstem astrocytes between control and PA-treated groups. ( B ): bar graphs showing fluorescence intensity of p16 INK4a between the control and PA-treated groups. Data are means ± SEM ( n = 4 replicates per group). Scale bar indicates 100 μm. * significant difference ( p < 0.05) between the groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Immunofluorescence, Staining, Marker, Control, Fluorescence

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: Immunofluorescence staining of p21CIP1 in the HBAs. ( A ): immunofluorescence staining for the senescence marker p21CIP1 in the human brainstem astrocytes between control and PA-treated groups. ( B ): bar graphs showing fluorescence intensity of p21CIP1 between the control and PA-treated groups. Data are means ± SEM ( n = 4 replicates per group). Scale bar indicates 100 μm. * significant difference ( p < 0.05) between the groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Immunofluorescence, Staining, Marker, Control, Fluorescence

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: PA-induced senescence causes SASP factor production in the HBAs. ( A – J ): changes in the mRNA levels of various proinflammatory SASP factors in the human brainstem astrocytes between control and PA-treated groups. Data are means ± SEM ( n = 5–6 replicates per group). * significant difference ( p < 0.05) between the groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Control

Journal: Nutrients

Article Title: Palmitic Acid Induces Oxidative Stress and Senescence in Human Brainstem Astrocytes, Downregulating Glutamate Reuptake Transporters—Implications for Obesity-Related Sympathoexcitation

doi: 10.3390/nu16172852

Figure Lengend Snippet: PA treatment downregulates glutamate reuptake transporter expression in the HBAs. ( A , B ): changes in the mRNA levels glutamate re-uptake transporters, EAAT1 and EAAT2 in the human brainstem astrocytes between control and PA-treated groups. Data are means ± SEM ( n = 4–6 replicates per group). * significant difference ( p < 0.05) between the groups.

Article Snippet: Human brainstem astrocytes (HBAs) purchased from ScienCell Research Laboratories were used in all the experiments.

Techniques: Expressing, Control