rabbit polyclonal anti cd133  (Proteintech)

Journal: Cell Death & Disease

Article Title: Temozolomide promotes glioblastoma stemness expression through senescence-associated reprogramming via HIF1α/HIF2α regulation

doi: 10.1038/s41419-025-07617-w

Figure Lengend Snippet: A Immunofluorescence analysis showed high expression levels of stemness markers CD133, Nestin, and transcription factors Sox2 and Klf4 in aggregation cells. B Elevated expression levels of CD133, Nestin, Sox2, and Klf4 were observed after two weeks of culturing aggregation cells. C Western blot analysis confirmed increased expression of stemness markers CD133, CD15, Nestin, Sox2, and Klf4 after one to two weeks of TMZ treatment, with higher expression levels in aggregation cells. D Aggregation cells (group ②) demonstrated greater invasive capacity compared to controls (group ①). Co-culturing control and aggregation cells showed a higher number of migrating cells when control cells were co-cultured with aggregation cells (group ④) than with control cells (group ③). E RFP-labeled aggregation cells co-cultured with aggregation cells exhibited the highest invasive capacity (group ④) compared to other groups, in the order: group ④ > group ③ (RFP-aggregation cells co-cultured with control cells) > group ② (RFP-control cells co-cultured with aggregation cells) > group ① (RFP-control cells co-cultured with control cells). * P < 0.05 was determined using Student’s t -test.

Article Snippet: These cells were incubated with polyclonal rabbit anti-human CD133 + IgGs (Miltenyi Biotech, Germany) at 4 °C for 15 min, washed with PBS containing 1% BSA, and resuspended in PBSE (108 cells/300 μl).

Techniques: Immunofluorescence, Expressing, Western Blot, Control, Cell Culture, Labeling

Journal: Cell Death & Disease

Article Title: Temozolomide promotes glioblastoma stemness expression through senescence-associated reprogramming via HIF1α/HIF2α regulation

doi: 10.1038/s41419-025-07617-w

Figure Lengend Snippet: A GSEA analysis indicated upregulation of senescence-associated hallmarks and inhibition of growth-promoting pathways. B The heatmap of U87 overlapping differentially expressed genes (DEGs) with SASP at the mRNA level revealed upregulation of most DEGs, including IL6, IL7, CXCL3, CXCL2, ICAM1, CCL2, CCL3, MMP7, and TIMP1. Conversely, senescence-inhibiting genes such as CDC25B, CDC25C, CDC25A, CDKN2D, MSH6, and MSH5 were downregulated. C RT-qPCR analysis demonstrated significant time-dependent increases in the expression of senescence-promoting genes, including IL1a, IL1b, IL6, IL8, CCL2, CDKN1A, CDKN2B, P53, and CXCL3, while the expression of the senescence-inhibiting gene MSH2 decreased significantly. D Mass spectrometry revealed high expression of senescence-promoting SASP proteins, including ITGA4, MMP15, FN1, IGFB3, and FAS, with a concomitant decrease in senescence-suppressing SASP proteins, MSH2 and MSH6. E ELISA confirmed increased expression of IL1a, IL6, and IL8 in a time-dependent manner following TMZ treatment. F SA-β-Gal staining revealed a significant time-dependent increase in β-Gal-positive cells, and the rate of β-Gal positivity was lower in aggregation cells compared to control cells under TMZ treatment. G C 12 FDG expression increased approximately threefold after one week of TMZ treatment in CD133 − CD15 − GBM cells, with lower levels of expression in aggregation cells under equivalent TMZ concentrations. H C 12 FDG-negative and -positive cells were cultured under TMZ for 21 days, showing significantly higher levels of CD133 and CD15 in C 12 FDG-positive cells over time, while CD133 and CD15 expression was not significant difference in C 12 FDG-negative cells. * P < 0.05 and # P > 0.05 were determined using Student’s t -test.

Article Snippet: These cells were incubated with polyclonal rabbit anti-human CD133 + IgGs (Miltenyi Biotech, Germany) at 4 °C for 15 min, washed with PBS containing 1% BSA, and resuspended in PBSE (108 cells/300 μl).

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Mass Spectrometry, Enzyme-linked Immunosorbent Assay, Staining, Control, Cell Culture

Journal: Cell Death & Disease

Article Title: Temozolomide promotes glioblastoma stemness expression through senescence-associated reprogramming via HIF1α/HIF2α regulation

doi: 10.1038/s41419-025-07617-w

Figure Lengend Snippet: A GSEA analysis demonstrated significant upregulation of hypoxia hallmark pathways in newly formed aggregation cells. B HIF1α and HIF2α knockout CD133 − CD15 − cells cultured under TMZ for two months exhibited the lowest expression levels of stemness markers CD133, CD15, Nestin, and transcription factors Sox2 and Klf4. Cells with simultaneous knockout showed the least expression, followed by single knockouts, compared to the control. C HIF1α and HIF2α knockout CD133 − CD15 − cells cultured with TMZ for two months exhibited almost no aggregation formation. Aggregation formation was significantly reduced in single knockouts compared to the control. D Apoptosis and necrosis rates were most significantly increased in cells with simultaneous HIF1α and HIF2α knockouts, followed by single knockouts, compared to the control. E Cell cycle analysis revealed that control cells arrested in the G2/M phase, while HIF1α and HIF2α knockout cells entered the S phase. F The rates of β-Gal-positive and C 12 FDG-positive cells decreased most significantly in simultaneous HIF1α and HIF2α knockout cells, with intermediate decreases in single knockouts compared to the control. G RT-qPCR analysis showed the lowest expression of SASP factors, including IL1a, IL1b, IL6, IL8, CCL2, and others, in simultaneous knockouts, followed by single knockouts, compared to the control in U118 CD133 − CD15 − cells. H GO analysis of miRNA sequences from simultaneous and single HIF1α and HIF2α knockouts revealed activation of terms associated with senescence and stemness, such as stem cell population maintenance, DNA replication, cell cycle arrest, centrosomes, and p53 binding. I KEGG pathway analysis of miRNA sequences from simultaneous and single HIF1α and HIF2α knockouts indicated activation of pathways associated with senescence and stemness, including cell cycle regulation, cellular senescence, Wnt signaling, regulation of pluripotency in stem cells, focal adhesion, and autophagy. * P < 0.05, ** P < 0.01, and # P > 0.05 were determined using Student’s t -test.

Article Snippet: These cells were incubated with polyclonal rabbit anti-human CD133 + IgGs (Miltenyi Biotech, Germany) at 4 °C for 15 min, washed with PBS containing 1% BSA, and resuspended in PBSE (108 cells/300 μl).

Techniques: Knock-Out, Cell Culture, Expressing, Control, Cell Cycle Assay, Quantitative RT-PCR, Activation Assay, Binding Assay

Journal: The Journal of Physiological Sciences : JPS

Article Title: Changes in adrenoceptor expression level contribute to the cellular plasticity of glioblastoma cells

doi: 10.1016/j.jphyss.2025.100016

Figure Lengend Snippet: The antibodies used for Western blotting.

Article Snippet: CD133 rabbit polyclonal , 1:2000 , Proteintech , 18470 −1-AP.

Techniques: Western Blot, Produced

Journal: iScience

Article Title: CXCL3/TGF-β-mediated crosstalk between CAFs and tumor cells augments RCC progression and sunitinib resistance

doi: 10.1016/j.isci.2024.110224

Figure Lengend Snippet: CAFs-CM and CXCL3 regulated ERK1/2 signaling pathway (A and B) Western blot analysis and quantification of ERK1/2 and phosphorylated-ERK1/2 expression in Renca cells stimulated with CAFs-CM or CXCL3. (C) Expression of ERK1/2 and phosphorylated-ERK1/2 in Renca cells stimulated with conditioned medium from CAFs with CXCL3 interference. (D) Protein expression of ERK1/2 and phosphorylated-ERK1/2 in tumor tissue of tumor-bearing mice. (E and F) Protein expression of N-cadherin and CD133 in Renca cells stimulated with CAFs-CM or CXCL3. (G) Protein expression of CD133 and E-cadherin in tumor tissue of tumor-bearing mice. Data are shown as mean ± SEM, n = 3∼6, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Rabbit anti-CD133 Polyclonal Antibody , Proteintech , Cat# 18470-1-AP; RRID:AB_2172859.

Techniques: Western Blot, Expressing

Journal: iScience

Article Title: CXCL3/TGF-β-mediated crosstalk between CAFs and tumor cells augments RCC progression and sunitinib resistance

doi: 10.1016/j.isci.2024.110224

Figure Lengend Snippet:

Article Snippet: Rabbit anti-CD133 Polyclonal Antibody , Proteintech , Cat# 18470-1-AP; RRID:AB_2172859.

Techniques: Recombinant, Cell Culture, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, RNA Extraction, Sequencing, Over Expression, Plasmid Preparation, Software