CD44 siRNA oligonucleotides were purchased from Sheng Gong (Shanghai, China). siRNA transfection was performed using transfection reagent (Yeasen, 40806ES02) according to the manufacturer’s instructions. Knockdown efficiency was demonstrated by RT-PCR 2 days after transfection.
Transfection reagent
Manufactured by Yeasen
Sourced in China
Transfection reagent is a laboratory product used to facilitate the introduction of genetic material, such as DNA or RNA, into cells. It serves as a delivery vehicle to enable the uptake of these molecules by the target cells, allowing for the study of gene expression, protein function, and other cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Confocal microscopy, by Leica (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Gfp lc3b, by Addgene (1 mentions)
Puromycin, by Beyotime (1 mentions)
Doxycycline, by Beyotime (1 mentions)
Trieasy total rna extraction reagent, by Yeasen (1 mentions)
Anti vegf, by Proteintech (1 mentions)
Senescence associated β galactosidase sa β gal kit, by Beyotime (1 mentions)
Annexin 5 fitc, by BioLegend (1 mentions)
Complementary dna synthesis supermix, by Yeasen (1 mentions)
Dutasteride, by Apexbio (1 mentions)
Goat anti rabbit igg h l hrp, by Fcmacs (1 mentions)
Anti nuclear factor κb nf κb p65, by Cell Signaling Technology (1 mentions)
Rnase a, by Yeasen (1 mentions)
Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Anti β actin, by Proteintech (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Solarbio (1 mentions)
Sybr green master mix, by Yeasen (1 mentions)
4 protocols using transfection reagent
1
CD44 Knockdown Using siRNA
2
GFP-LC3 Transfection and Autophagy Monitoring
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Cells adhered
to slides were transfected
with GFP-LC3 plasmid using the transfection reagent (Yeasen), according
to the manufacturer’s protocol. After transfection for 24 h,
the cells were treated with C21 fraction, alone or with CQ, and then
incubated for 24 h. Finally, the cells were stained with DAPI and
an anti-quenching agent (1:1000, Beyotime) and observed using confocal
microscopy (Leica).
to slides were transfected
with GFP-LC3 plasmid using the transfection reagent (Yeasen), according
to the manufacturer’s protocol. After transfection for 24 h,
the cells were treated with C21 fraction, alone or with CQ, and then
incubated for 24 h. Finally, the cells were stained with DAPI and
an anti-quenching agent (1:1000, Beyotime) and observed using confocal
microscopy (Leica).
3
SARS-CoV-2 ORF3a Overexpression and Autophagy Tracking
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SARS-CoV-2 ORF3a (Gene ID: 43740569) with HA-Tag was cloned into the pCAGGS through restriction sites EcoRI and BamHI. Plasmids GFP-mCherry-LC3B (#123230) and GFP-LC3B (#11546) were purchased from Addgene (Watertown, MA, USA). Transfection reagent (Yeasen, Shanghai, China) was used according to the manufacturer’s protocol.
4
Antibody-based Cellular Pathway Analysis
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The following antibodies were used: anti-SRD5A1 (Proteintech, #66329-1-Ig, 1:1,000 dilution), anti-nuclear factor-κB (NF-κB) p65 (Cell Signaling Technology, #8242, 1:1,000 dilution), anti-Phospho-NF-κB p65 (Cell Signaling Technology, #3033, 1:1,000 dilution), anti-VEGF (Proteintech, #19003-1-AP, 1:1,000 dilution), and anti-β-actin (Proteintech, #66009-1-Ig, 1:5,000 dilution). Secondary antibodies included goat anti-Rabbit IgG(H+L) HRP (Fcmacs, #FMS-Rb01, 1:5,000 dilution) or mouse (Affinity, #S0002, 1:10,000 dilution). 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) was purchased from Solarbio (Beijing, China), dutasteride was purchased from APExBIO (Houston, TX, USA), propidium iodide (PI) was purchased from XunBei (Nanjing, China), and Annexin-V-FITC was purchased from Biolegend (San Diego, CA, USA). Pierce TM BCA Protein Assay kit, transfection reagent, and RNase A were purchased from YEASEN (Shanghai, China). Senescence-associated β-galactosidase (SA-β-gal) kit, puromycin, and doxycycline were purchased from Beyotime Institute of Biotechnology (Shanghai, China). TRIeasy™ Total RNA Extraction Reagent, complementary DNA synthesis superMix, and SYBR Green Master Mix were purchased from YEASEN (Shanghai, China).
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