PC3/Doc cells were transiently transfected with dominant‐negative PCMV5‐AKT1‐K179M (AKT1‐DN), ATF3 siRNA (described previously 13 ) or HDAC5 siRNA (sc‐35542) (Santa Cruz Biotechnology) using lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). Empty vectors PCMV5 served as controls. After 24‐hrs transfection, cells were treated with TSA or vehicle for an additional 24 hrs, and cell lysates were subjected to a Western blot assay. Cell viability and death were determined by MTT assays.
Atf3 sirna
Manufactured by Santa Cruz Biotechnology
Sourced in United States
ATF3 siRNA is a small interfering RNA (siRNA) product designed to target the ATF3 gene. The ATF3 gene encodes a transcription factor that plays a role in various cellular processes. This siRNA product can be used to downregulate the expression of the ATF3 gene, potentially providing a tool for researchers studying the functions and mechanisms of ATF3 in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco s modified eagle medium dmem f 12 1 1 modified medium dmem f 12, by Lonza (3 mentions)
β actin, by Cell Signaling Technology (3 mentions)
Poly adp ribose polymerase parp, by Cell Signaling Technology (2 mentions)
Dmem f 12 1 1 modified medium dmem f 12, by Lonza (2 mentions)
3 4 5 dimethylthizaol 2 yl 2 5 diphenyl tetrazolium bromide mtt, by Merck Group (2 mentions)
Atf3 antibody, by Santa Cruz Biotechnology (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Naringenin nar, by Merck Group (1 mentions)
Kahweol, by Merck Group (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
Total erk1 2, by Cell Signaling Technology (1 mentions)
Total p38, by Cell Signaling Technology (1 mentions)
Total jnk, by Cell Signaling Technology (1 mentions)
Total ikkα, by Cell Signaling Technology (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
Total gsk3β, by Cell Signaling Technology (1 mentions)
P ikkα, by Cell Signaling Technology (1 mentions)
Sb216763, by Merck Group (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
P erk1 2, by Cell Signaling Technology (1 mentions)
8 protocols using atf3 sirna
1
Regulating AKT1, ATF3, and HDAC5 in PC3 cells
2
Naringenin Modulates Stress Pathways
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Naringenin (NAR) was purchased from Sigma Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle medium (DMEM)/F-12 1:1 Modified medium (DMEM/F-12) was purchased from Lonza (Walkersville, MD, USA). Antibodies against ATF3, Poly ADP ribose polymerase (PARP) and β-actin were purchased from Cell Signaling (Beverly, MA, USA). PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor), SB216763 (GSK3β inhibitor) and BAY11-7082 (NF-κB inhibitor) were purchased from Calbiochem (San Diego, CA, USA). ATF3 siRNA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). ATF3 constructs used in this study were kindly provided from Dr. Seong Ho Lee (University of Maryland, College Park, MD, USA). All chemicals were purchased from Fisher Scientific, unless otherwise specified.
3
Kahweol Regulation of ATF3 Signaling
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Kahweol was purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle medium (DMEM)/F-12 1:1 Modified medium (DMEM/F-12) was purchased from Lonza (Walkersville, MD, USA). Antibodies against ATF3, Poly ADP ribose polymerase (PARP) and β-actin were purchased from Cell Signaling (Beverly, MA, USA). PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor) and LiCl (GSK3β inhibitor) were purchased from Calbiochem (San Diego, CA, USA). ATF3 siRNA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). ATF3 promoter constructs used in this study were kindly provided by Dr. Seong Ho Lee (University of Maryland, College Park, MD, USA). All chemicals were purchased from Fisher Scientific (Hampton, NH, USA), unless otherwise specified.
4
Cellular Signaling Pathway Assay
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Dulbecco’s Modified Eagle medium (DMEM)/F-12 1:1 Modified medium (DMEM/F-12) was purchased from Lonza (Walkersville, MD, USA). Antibodies against ATF3, poly (ADP-ribose) polymerase (PARP), p-ERK1/2, total-ERK1/2, p-p38, total-p38, p-JNK, total-JNK, p-GSK3β, total-GSK3β, p-IKKα, total-IKKα, and β-actin were purchased from Cell Signaling (Beverly, MA, USA). PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor), LiCl (GSK3β inhibitor), and BAY11-7082 (IKKα inhibitor) were purchased from Calbiochem (San Diego, CA, USA). ATF3 siRNA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). ATF3 promoter constructs used in this study were kindly provided by Seong Ho Lee (University of Maryland, College Park, MD, USA). All chemicals were purchased from Fisher Scientific, unless otherwise specified.
5
Investigating ATF3 Regulation in Cell Signaling
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Cell culture media, Dulbecco's Modified Eagle medium (DMEM)/F-12 1:1 Modified medium (DMEM/F-12) was purchased from Lonza (Walkersville, MD, USA). 3-(4,5-dimethylthizaol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) were purchased from Sigma Aldrich (St. Louis, MO, USA). SB203580, PD98059 were purchased from Calbiochem (San Diego, CA, USA). SB216763 were purchased from Sigma Aldrich. ATF3 antibody and ATF3 siRNA were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Antibodies against β-actin, Poly (ADP-ribose) polymerase (PARP), p38, p-p38, ERK1/2, p-ERK1/2, p-GKS3β and GKS3β were purchased from Cell Signaling (Bervely, MA, USA). ATF3 promoter constructs (-1420/+34, -718/+34, -514/+34, -318/+34, -147/+34 and -84/+34, pATF3-514 del Ftz and pATF3-514 del CRE) were kindly provided by Dr. S-H Lee (University of Maryland College Park, Maryland, USA). All chemicals were purchased from Fisher Scientific, unless otherwise specified.
6
Evaluation of ATF3 Regulation in Cell Culture
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Cell culture media, Dulbecco’s Modified Eagle medium (DMEM)/F-12 1:1 Modified medium (DMEM/F-12) was purchased from Lonza (Walkersville, MD, USA). The 3-(4,5-dimethylthizaol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and SP600125 was purchased from Sigma Aldrich (St. Louis, MO, USA). SB203580 and PD98059 were purchased from Calbiochem (San Diego, CA, USA). ATF3 antibody and ATF3 siRNA were provided from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Antibodies against β-actin and poly (ADP-ribose) polymerase (PARP), and control siRNA were purchased from Cell Signaling (Bervely, MA, USA). ATF3 promoter constructs (-1420/+34, -718/+34, -514/+34, -318/+34, -147/+34 and -84/+34) were kindly provided by Dr. S-H Lee (University of Maryland College Park, MD, USA). All chemicals were purchased from Fisher Scientific, unless otherwise specified.
7
ATF3 Silencing in Cell Migration
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Small interfering RNA interference nucleotide sequence of ATF3 (ATF3-siRNA) was designed (Santa Cruz Biotech Co), and an independent nucleic sequence with the same base number was synthesized as negative control (NC-siRNA). Liposome transfection was in accordance with Lipofectamine TM 3000 (Invitrogen) with 100 nM of NC-SiRNA or ATF3-siRNA according to the manufacturer’s instructions. Transfection efficiency was evaluated by ATF3 mRNA and protein level at 24 hours after liposome transfection. Subsequently, the transfected cells were evaluated by wound healing, Transwell migration, and Transwell invasion assays.
8
Modulation of LPS-induced inflammation
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Raw 264.7 cells were maintained in DMEM medium containing 10% fetal bovine serum, and 5×105 cells/well in 12-well plates were transfected with α1-AMPK siRNA, ATF-3 siRNA, or negative control siRNA (Santa Cruz, CA) (100 nM/well) using 6 µl of Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA) in OPTI MEM medium for 6 hr. Medium was then replaced with DMEM containing 10% FBS and 18 hr later, cells were treated with berberine (10 and 20 µM) for 20 hr and then with 10 ng/ml LPS for 30 min (western blot of phosphoprotein) and for 4 hr (ELISA). Efficiencies of ATF-3 and AMPK knockdowns were determined by Western blotting and by RT-PCR.
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