Journal: bioRxiv

Article Title: Clusterin can mediate apoptosis-induced molecular mechanisms in immune thrombocytopenia

doi: 10.1101/2023.09.26.559483

Figure Lengend Snippet: imRNA expression levels of Total Clusterin, Clusterin 1 and Clusterin 2 in platelets from our ITP patient groups. Total RNA from platelets was isolated and subjected to RT-qPCR with 18S rRNA as internal control. Relative mRNA levels of each sample normalized to healthy controls are shown as means (n=9) ± SD. Error bars show SD. Statistical analyses were performed using one-way ANOVA followed by multiple comparisons tests to compare the mean ranks between the groups. Means are significantly different with < 0.001 (highly significant) (B) Clusterin expression was assessed by Western blotting in healthy controls (lane1), newly diagnosed (ND) ITP (lane 2) and chronic ITP patients (lane 3). A representative immunoblotting image was selected (N=3). .

Article Snippet: MEG-01 cells were incubated with siRNAs gene against Clusterin and negative control siRNA (Santa Cruz Biotechnology) for 48 hours according to manufacturer’s instructions.

Techniques: Expressing, Isolation, Quantitative RT-PCR, Western Blot

Journal: bioRxiv

Article Title: Clusterin can mediate apoptosis-induced molecular mechanisms in immune thrombocytopenia

doi: 10.1101/2023.09.26.559483

Figure Lengend Snippet: mRNA levels of Total Clusterin and Clusterin 1 in PBMCs from our ITP patient groups. Total RNA was isolated and subjected to RT-qPCR with 18S rRNA as internal control. Relative mRNA levels of each sample normalized to healthy controls are shown as means (n=2) ± SD. Error bars show SD. Statistical analyses were performed using one-way ANOVA followed by multiple comparisons tests to compare the mean ranks between the groups. Means are significantly different with p < 0.001 (highly significant).

Article Snippet: MEG-01 cells were incubated with siRNAs gene against Clusterin and negative control siRNA (Santa Cruz Biotechnology) for 48 hours according to manufacturer’s instructions.

Techniques: Isolation, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Clusterin can mediate apoptosis-induced molecular mechanisms in immune thrombocytopenia

doi: 10.1101/2023.09.26.559483

Figure Lengend Snippet: MK cell line MEG-01 treated with ITP plasma shows the different clusterin\ expression after siRNA transfections. mRNA levels of Clusterin determined by qRT-PCR in MEG-01 cell line treated with plasma from ITP patients compared to plasma from healthy Controls and trasfected with siRNA Clusterin. Statistical analyses were performed using one-way ANOVA followed by multiple comparisons tests to compare the mean ranks between groups. Expression levels are shown as means; error bars show SDs and P values are indicated.

Article Snippet: MEG-01 cells were incubated with siRNAs gene against Clusterin and negative control siRNA (Santa Cruz Biotechnology) for 48 hours according to manufacturer’s instructions.

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: Molecular Neurodegeneration

Article Title: BACE1 regulates expression of Clusterin in astrocytes for enhancing clearance of β-amyloid peptides

doi: 10.1186/s13024-023-00611-w

Figure Lengend Snippet: Clusterin underlies astrocyte endocytosis of Aβ in vitro. A Western blot of Bace1 -null and WT-primary astrocytes pretreated with either 80 pmol of Clu siRNA or 80 pmol of control scrambled siRNA and incubated with 2 μM of Aβ 42 for indicated times. Images indicate major bands for Aβ 42 (human, oligomeric) and GAPDH. B Quantification of Western blot Aβ 42 band intensity normalized to actin ( N = 3, * p -value < 0.05, ** p -value < 0.01, *** p -value < 0.001; One-way ANOVA with Tukey post testing comparing between samples. C Confocal imaging of astrocyte primary cultures from BACE1-null and WT and pretreated with either Clu or control scrambled siRNA then treated with 2 μM of Aβ 42 tagged with fluorescent Hilyte-555 for indicated incubation times. Stained with phalloidin (green) to mark F-actin and ToPro3 (blue) to mark nuclei. Scale bars represents 10 μm

Article Snippet: Knockdown of Clu in astrocyte cultures was attained by using Clusterin siRNA reagents (Santa Cruz Biotechnology, catalog no. sc-43689) or control scrambled siRNA (Santa Cruz Biotechnology, catalog no. sc-37007) according to manufacturer’s instructions.

Techniques: In Vitro, Western Blot, Incubation, Imaging, Staining

Journal: Molecular Neurodegeneration

Article Title: BACE1 regulates expression of Clusterin in astrocytes for enhancing clearance of β-amyloid peptides

doi: 10.1186/s13024-023-00611-w

Figure Lengend Snippet: Differentially expressed genes in Bace1-deleted astrocytes

Article Snippet: Knockdown of Clu in astrocyte cultures was attained by using Clusterin siRNA reagents (Santa Cruz Biotechnology, catalog no. sc-43689) or control scrambled siRNA (Santa Cruz Biotechnology, catalog no. sc-37007) according to manufacturer’s instructions.

Techniques: Binding Assay

Journal: Molecular Neurodegeneration

Article Title: BACE1 regulates expression of Clusterin in astrocytes for enhancing clearance of β-amyloid peptides

doi: 10.1186/s13024-023-00611-w

Figure Lengend Snippet:

Article Snippet: Knockdown of Clu in astrocyte cultures was attained by using Clusterin siRNA reagents (Santa Cruz Biotechnology, catalog no. sc-43689) or control scrambled siRNA (Santa Cruz Biotechnology, catalog no. sc-37007) according to manufacturer’s instructions.

Techniques: