human clu clusterin elisa kit  (Elabscience Biotechnology)

Journal: Molecular Neurodegeneration

Article Title: Alzheimer’s disease protective allele of Clusterin modulates neuronal excitability through lipid-droplet-mediated neuron-glia communication

doi: 10.1186/s13024-025-00840-1

Figure Lengend Snippet: AD risk SNP rs1532278 shows allele-specific open chromatin (ASoC) and cis-regulates CLU expression in iGlut. A Schematic research design. ATAC-seq was performed to identify OCRs and GWAS risk SNPs that showed ASoC in iPSC Glutaminergic (iGlut), GABAergic (iGABA), and dopaminergic (iDA) neurons, and microglia (iMG) and astrocytes (iAst), which is followed by transcription factor (TF) binding prediction to confirm putative functional AD risk SNP, CRIPSR-cas9 SNP editing, and functional assays in cells. B ASoC mapping identifies intronic rs1532278 as a putatively functional SNP among several other GWAS risk SNPs equivalently associated with AD at the CLU locus. Note that rs1532278 is the only SNP within the neuronal OCR with a stronger OCR peak in iGlut. C T and C alleles of rs1532278 show differential allelic ATAC-seq reads (i.e., ASoC) in iGlut. The bottom panel shows the two most conserved TF binding motifs at the SNP site. D Schematic CLU gene structure near rs1532278 (upper panel) and a diagram showing CRISPR-Cas9 editing of rs1532278 in two iPSC lines (CD05 and CD07; T/C) to covert T/C lines to isogenic T/T and C/C lines (middle panel, representative Sanger sequencing result from CD07 line). Representative images of iGlut (CD07 line) of all three genotypes are also shown (bottom panel); MAP2 and HuNu (human nuclear antigen) staining shows the morphology of iGlut and neuron purity in iGlut-mAst co-cultures. E – F ISL2 ChIP-qPCR on day 30 iGlut-mAst co-cultures. n = 3 biological replicates from one clone per line of one independent differentiation. G - H ISL2 siRNA knockdown in day- 30 pure iGlut (T/T) cultures. Samples of 72 h post-siRNA transfection were used for qPCR. n = 3 biological replicates from one clone per line of one independent differentiation. I Neuronal CLU mRNA level in isogenic iGlut-mAst co-cultures of different genotype of rs1532278 (human-specific CLU qPCR assay was used). n = 6 biological replicates per group (2–3 clones per line and 2–3 biological replicates for each clone) from two independent differentiations of each line. J Secreted CLU (sCLU) detected by ELISA from the supernatant of iGlut-mAst co-cultures. n = 4 biological replicates per group (2 clones per line and 2 biological replicates per clone) from two independent differentiations of each line. K - L Immunofluorescence staining of CLU in day 25 pure iGlut cultures. n = 6–7 coverslips per group (In total: 2 clones per line and 3–4 coverslips for each clone and 3–5 images per coverslip; shown are example images of CD05) from two independent differentiations of each line. Data, mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. Scale bars are indicated in each image

Article Snippet: ELISA quantifications of human CLU (R&D system, DCLU00, specific for human CLU), human Aβ 1–40 (R&D system, DAB140B), human Aβ 1–42 (R&D system, DAB142), and mouse CLU (R&D system, MCLU00, specific for mouse CLU) were performed according to the vendors’ protocol.

Techniques: Expressing, Binding Assay, Functional Assay, CRISPR, Sequencing, Staining, ChIP-qPCR, Knockdown, Transfection, Clone Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Human reproduction (Oxford, England)

Article Title: CDC42 deficiency leads to endometrial stromal cell senescence in recurrent implantation failure.

doi: 10.1093/humrep/deae246

Figure Lengend Snippet: Figure 1. Cellular senescence in the stroma of recurrent implantation (RIF) failure endometrium. (A) Gene ontology (GO) biological process enrichment of the differentially expressed genes (DEGs) between fertile control (CTR) patients and patients with RIF in SRP224538. (B) FPKM level of TP53 in CTR patients and RIF patients in SRP224538. (C) FPKM level of IL6 in CTR patients and RIF patients in SRP224538. (D–F) Levels of IL6, CLU, and sST2 secretion in primary endometrial stromal cell supernatant from CTR and RIF patients before and after 72 h of 8Br-cAMPþMPA treatment. (G and H) Representative images of immunohistochemical (IHC) staining of P16 in mid-secretory endometrium of CTR patients (n ¼ 22) versus RIF patients (n ¼ 22). Black arrows indicate typical P16þ cells. Quantitative analysis by number of P16þ cells per 200× field for P16 staining. (I and J) Representative images of IHC staining of P21 in mid-secretory endometrium of CTR patients (n ¼ 22) versus RIF patients (n ¼ 22). Black arrows indicate typical P21þ

Article Snippet: Commercially available ELISA kits were utilized to measure the level of secreted protein (PRL: Elabscience, No. E-EL-H0141; IGFBP1: Elabscience, No. E-EL-H0442; IL6: Elabscience, No. E-EL-H6156; IL8: MULTI SCIENCES, No. EK10896; CLU: Elabscience, No. E-EL-H0038; sST2: Elabscience, No. E-EL-H6082) in collected supernatants according to the manufacturer’s instructions.

Techniques: Control, Immunohistochemical staining, Immunohistochemistry, Staining

Journal: Human reproduction (Oxford, England)

Article Title: CDC42 deficiency leads to endometrial stromal cell senescence in recurrent implantation failure.

doi: 10.1093/humrep/deae246

Figure Lengend Snippet: Figure 2. CDC42 knockdown accelerates senescence of differentiated endometrial stromal cells (EnSCs). (A–C) Expression of CDC42, CDKN2A, and CDKN1A mRNA levels in primary EnSCs after 72 h of adenovirus treatment. (D) Expression of CDC42, P21, P53, and p-γ-H2AX protein levels in primary EnSCs after 72 h of adenovirus treatment. (E) Expressions of collagen I, collagen III, collagen IV, and MMP2 protein levels in primary EnSC after 72 h of adenovirus treatment. (F–H) Expressions of CDC42, CDKN2A, and CDKN1A mRNA levels in primary EnSC transfected with Ad-GFP or Ad-shCDC42-GFP following with 72 h of 8Br-cAMPþMPA treatment. (I–K) Level of IL6, CLU, and sST2 secretion in primary EnSCs transfected with Ad-GFP or Ad-shCDC42- GFP following with 72 h of 8Br-cAMPþMPA treatment. (L) Expression of CDC42, P21, and P53 protein levels in primary EnSCs transfected with Ad-GFP or Ad-shCDC42-GFP following with 72 h of 8Br-cAMPþMPA treatment. (M and N) Representative images of P16 and P21 immunofluorescence staining in primary EnSCs transfected with Ad-GFP or Ad-shCDC42-GFP following with 72 h of 8Br-cAMPþMPA treatment. (O and P) SA-β-gal staining of primary EnSCs after 48 h of Ad-GFP or Ad-shCDC42-GFP adenovirus treatment and then different days of 8Br-cAMPþMPA treatment. Quantitative analysis of integrated optical density for SA-β-gal staining, with value of the Ad-GFP group at 0 h set as 1. Mean ± SEM. P < 0.05, P < 0.01, P<0.001. ANOVA with Tukey’s multiple comparisons test. Two-way ANOVA with the Bonferroni multiple comparisons test in (P).

Article Snippet: Commercially available ELISA kits were utilized to measure the level of secreted protein (PRL: Elabscience, No. E-EL-H0141; IGFBP1: Elabscience, No. E-EL-H0442; IL6: Elabscience, No. E-EL-H6156; IL8: MULTI SCIENCES, No. EK10896; CLU: Elabscience, No. E-EL-H0038; sST2: Elabscience, No. E-EL-H6082) in collected supernatants according to the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Transfection, Immunofluorescence, Staining