quantitative cellular senescence assay kit (sa–β-gal, fluorometric) cba-232  (Cell Biolabs Inc)

Journal: Molecular Cell

Article Title: Deregulated Expression of Mammalian lncRNA through Loss of SPT6 Induces R-Loop Formation, Replication Stress, and Cellular Senescence

doi: 10.1016/j.molcel.2018.10.011

Figure Lengend Snippet: SPT6 Depletion Induces DNA Replication Stress and Cellular Senescence (A) Western blot of whole-cell extract using indicated antibodies, following HeLa cell SPT6 depletion. EXOSC3 is loading control. (B) Microscopy of β-galactosidase staining cells (blue) and associated quantification of β-galactosidase staining cells with SPT6 or INTS11 depletions. Blue arrowhead identifies β-gal positive cells. (C) Western blot with indicated antibodies of engineered HeLa cells ± Tet-inducible V5-RNase H1 overexpression. After 4 hr siRNA transfection, fresh media ±1 μg/mL tetracycline (Tet) was added and incubated for 16 hr. (D) DIP-qPCR analysis of YWH2 pre-mRNA and PROMPT ± Tet induction of V5 RNase H1. Data are represented as mean ± SEM. (E) Western blot with indicated antibodies of whole-cell extract from HeLa cells ± SPT6, +/− following 18 hr aphidicolin (APH) treatment. (F) qRT-PCR of YWHAZ chromatin-bound RNA from cells as in (E). Data are represented as mean ± SEM. (G) FACS of HeLa cells (+/− SPT6 depletion). After 6 hr with siRNA DMSO (control) or APH was added for 18 hr. Quantitation shown on the right is mean ± SEM. (H) Quantification of β-galactosidase-stained cells treated with DMSO or APH. Top: siRNA transfection (6 hr) and then DMSO or APH added and incubated (18 hr). Total was 24 hr. Bottom: siRNA transfection (24 hr) and then DMSO or APH added and incubated (24 hr). Total was 48 hr. (I) Left: meta-analysis of mNET-seq/total CTD in –/+ 2-kb regions from HeLa DNA replication origins (1 kb). Right: boxplots quantifying mNET-seq/total CTD signals over HeLa DNA replication origins. (J) LincRNA LOC646626 gene loci (+ strand) as example of mNET-seq and ChrRNA-seq profiles over DNA replication origin. Magnified field of replication origin shown below. (K) Model: collision between Pol II and DNA replisome in SPT6-depleted cell. Pol II and DNA replisome are separated in wild-type cells. In SPT6-depleted cells, induced and extended lncRNA transcription in intergenic region causes deregulated Pol II collision with DNA replisome. R-loops may also cause DNA damage and cellular senescence. See also Figure S6 and and .

Article Snippet: Quantitative cellular senescence assay kit (SA- β-gal, Fluorometric) , Cell Biolabs , Cat# CBA-232.

Techniques: Western Blot, Control, Microscopy, Staining, Over Expression, Transfection, Incubation, Quantitative RT-PCR, Quantitation Assay

Journal: Molecular Cell

Article Title: Deregulated Expression of Mammalian lncRNA through Loss of SPT6 Induces R-Loop Formation, Replication Stress, and Cellular Senescence

doi: 10.1016/j.molcel.2018.10.011

Figure Lengend Snippet:

Article Snippet: Quantitative cellular senescence assay kit (SA- β-gal, Fluorometric) , Cell Biolabs , Cat# CBA-232.

Techniques: Positive Control, Sequencing, Purification, Staining, Software

Journal: Cell metabolism

Article Title: β-cell aging markers have heterogeneous distribution and are induced by insulin resistance

doi: 10.1016/j.cmet.2017.03.015

Figure Lengend Snippet: DATA AND SOFTWARE AVAILABILITY

Article Snippet: Cellular Senescence Live Cell Analysis Assay Kit (SA-β-gal, Fluorometric) , , Enzo Life Sciences , enz-kit 130-0010.

Techniques: Software, Immunostaining, Isolation, Recombinant, Microarray