Journal: bioRxiv

Article Title: Novel strategies of Raman imaging for monitoring intracellular retinoid metabolism in cancer cells

doi: 10.1101/2020.05.05.078410

Figure Lengend Snippet: Microscopy images of lipid droplets stained with Oil Red O dye in normal human astrocytes NHA (A) and high-grade glioblastoma U-87 MG (B and L) with bar plots of the total number of lipid droplets in NHA and U-87 MG , the area of LD per cell and the size of LDs were quantified by Image J software . The results represent the means +/- SEM of at least 6 representative cells (n(NHA)=6, n(U-87 MG)=9), Raman spectra of lipid droplets of normal astrocytes, profile I – TAG (orange), profile II – retinyl esters (blue) (C), microscopy image (D), Raman cluster image (E)(image size 50×50 μm, resolution 1 μm, integration time 0.3 second, number of Raman spectra 2500) and Raman image of the distribution of lipid droplets with profile I (F) and profile II (G) in NHA. Average Raman spectrum of lipid droplets obtained from cluster analysis (H), microscopy image (I), Raman cluster image (J)(image size 55×20 μm, resolution 1 μm, integration time 0.3 second, number of Raman spectra 1100) and fluorescence image of Oil Red O (K) and microscopy images of Oil Red O-stained lipid droplets (L) of U-87 MG high-grade glioblastoma. Raman analysis were performed at 532 nm laser excitation. Nucleus was labeled by red, lipid droplets – blue/orange, cytoplasm - green, mitochondria - magenta, cell border - light grey and area out of cell - dark grey.

Article Snippet: A normal human astrocyte (NHA) cell line (CC-2565; Lonza) and human glioblastoma (U-87 MG) cell line (HTB-14; ATCC) were used.

Techniques: Microscopy, Staining, Software, Fluorescence, Labeling

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Circ_0000189 Promotes the Malignancy of Glioma Cells via Regulating miR-192-5p-ZEB2 Axis

doi: 10.1155/2022/2521951

Figure Lengend Snippet: The expression characteristics of circ_0000189 in gliomas. (a) qRT-PCR was utilized to detect circ_0000189 expression in human glioma tissues and adjacent normal tissues (total = 50). (b) Circ_0000189 expression in human glioma cell lines (A172, U87, LN18, U251, LN229, and U118 cells), normal human astrocyte (NHA) cell line, and human normal brain glial cell line (HEB) was investigated by qRT-PCR. ∗∗ symbolizes P < 0.01, and ∗∗∗ symbolizes P < 0.001.

Article Snippet: Normal human astrocyte cell line (NHA), human normal brain glial cell line (HEB), and human glioma cell lines (A172, U87, LN18, U251, LN229, and U118 cells) were available from China Center for Type Culture Collection (CCTCC, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Circ_0000189 Promotes the Malignancy of Glioma Cells via Regulating miR-192-5p-ZEB2 Axis

doi: 10.1155/2022/2521951

Figure Lengend Snippet: miR-192-5p was a target of circ_0000189 in gliomas. (a) Bioinformatics predicted the binding site between circ_0000189 and miR-192-5p. (b) qRT-PCR was utilized to explore miR-192-5p expression in the normal human astrocyte (NHA) cell line and human glioma cell lines (A172, U87, LN18, U251, and LN229, as well as U118 cells). (c) Dual luciferase report assay was performed to verify the predicted binding site between miR-192-5p and circ_0000189. (d) RIP experiment was performed to verify the direct interaction between circ_0000189 and miR-192-5p. (e) After upregulating the expression of circ_0000189 in A172 cells and knocking down the expression of circ_0000189 in U118 cells, qRT-PCR was utilized to investigate miR-192-5p expression. (f) miR-192-5p expression in normal tissues and glioma tissues was detected by qRT-PCR. (g) Analysis of the correlation between circ_0000189 expressions and miR-192-5p expression in glioma tissues. ∗ symbolizes P < 0.05, ∗∗ symbolizes P < 0.01, and ∗∗∗ symbolizes P < 0.001.

Article Snippet: Normal human astrocyte cell line (NHA), human normal brain glial cell line (HEB), and human glioma cell lines (A172, U87, LN18, U251, LN229, and U118 cells) were available from China Center for Type Culture Collection (CCTCC, Wuhan, China).

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

Journal: International Journal of Molecular Sciences

Article Title: Allopregnanolone Promotes Migration and Invasion of Human Glioblastoma Cells through the Protein Tyrosine Kinase c-Src Activation

doi: 10.3390/ijms23094996

Figure Lengend Snippet: AKR1C1-4 are differentially expressed in GB cells. The expression of AKR1C1-4 was detected in a total protein extract from normal human astrocytes (HA) and different human GB cell lines. ( a ) Representative Western blots of AKR1C1-4 and α-Tubulin, which was used as a loading control. ( b ) Densitometric analysis graph. Each column represents the mean ± SEM. n = 3; * p < 0.05 U251, T98G, and LN229 vs. HA and U87 cell lines.

Article Snippet: For Western blot determination of AKR1C1-4 (37 kDa), 20 μg of normal human primary astrocytes lysate (HA; 1806, ScienCell, Carlsbad, CA, USA), and 20 μg of protein lysate of the cell lines were mixed with Laemmli 2X buffer (100 mM Tris-base pH 6.8, 20% glycerol, 4% SDS, 10% β-mercaptoethanol, and bromophenol blue) were boiled for 5 min and separated in a 12% SDS-PAGE gels at 80 V. The separated proteins were then transferred to nitrocellulose membranes (Millipore, Burlington, MA, USA) by electrophoresis in semi-dry conditions at 30 mA per membrane for 2 h. Membranes were blocked in agitation at 37 °C with a blocking solution (TBS buffer-0.1% Tween with 5% bovine serum albumin; InVitro, MEX) for 2 h; then, membranes were incubated with a mouse monoclonal AKR1C1-4 antibody (1:1000; sc-390419, Santa Cruz, CA, USA) overnight.

Techniques: Expressing, Western Blot, Control