transintro pl transfection reagent Search Results


transintro pl transfection reagent  (TransGen biotech co)
94
TransGen biotech co transintro pl transfection reagent
Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h <t>post-transfection,</t> the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.
Transintro Pl Transfection Reagent, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transintro+pl+transfection+reagent/pmc12172468-159-0-10?v=TransGen+biotech+co
Average 94 stars, based on 1 article reviews
transintro pl transfection reagent - by Bioz Stars, 2026-07
94/100 stars
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transintrotm el transfection reagent  (TransGen biotech co)
96
TransGen biotech co transintrotm el transfection reagent
Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h <t>post-transfection,</t> the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.
Transintrotm El Transfection Reagent, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transintro+pl+transfection+reagent/pm30336336-95-35-39?v=TransGen+biotech+co
Average 96 stars, based on 1 article reviews
transintrotm el transfection reagent - by Bioz Stars, 2026-07
96/100 stars
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transfection reagent  (TransGen biotech co)
94
TransGen biotech co transfection reagent
Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h <t>post-transfection,</t> the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.
Transfection Reagent, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transintro+pl+transfection+reagent/pmc11098806-231-15-18?v=TransGen+biotech+co
Average 94 stars, based on 1 article reviews
transfection reagent - by Bioz Stars, 2026-07
94/100 stars
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transintro pei transfection reagent  (TransGen biotech co)
93
TransGen biotech co transintro pei transfection reagent
Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h <t>post-transfection,</t> the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.
Transintro Pei Transfection Reagent, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transintro+pl+transfection+reagent/pmc12077208-291-28-32?v=TransGen+biotech+co
Average 93 stars, based on 1 article reviews
transintro pei transfection reagent - by Bioz Stars, 2026-07
93/100 stars
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transintro® el transfection reagent  (TransGen biotech co)
90
TransGen biotech co transintro® el transfection reagent
Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h <t>post-transfection,</t> the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.
Transintro® El Transfection Reagent, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transintro+pl+transfection+reagent/pm37376546-98-19-23?v=TransGen+biotech+co
Average 90 stars, based on 1 article reviews
transintro® el transfection reagent - by Bioz Stars, 2026-07
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transintro tm el transfection reagent  (Shanghai GenePharma)
90
Shanghai GenePharma transintro tm el transfection reagent
Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h <t>post-transfection,</t> the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.
Transintro Tm El Transfection Reagent, supplied by Shanghai GenePharma, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/transintro+pl+transfection+reagent/pmc11680124-78-16-5?v=Shanghai+GenePharma
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transintro tm el transfection reagent - by Bioz Stars, 2026-07
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transintro pl transfection reagent - 2 ×  (TransGen biotech co)
N/A
Transform your experiments with TransIntro™ PL Transfection Reagent! This powerful 2 × 0.75 mL reagent allows for efficient and reliable delivery of genetic material into cells. Take a daring step towards successful research with this
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Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Incubation, Isolation, Software

The KFERQ-like motif (QVDRL) in PEDV S protein is recognized and bound by HSC70. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-ΔCRR or Flag-tagged empty vector for 36 h. The proteins were immunoprecipitated in WCL using anti-Flag magnetic beads, and then the associated proteins were separated by 7.5% SDS-PAGE and stained with silver. The black arrow indicated the expressed S-ΔCRR, and the red arrow indicated a different immunoprecipitated protein band in the S-ΔCRR-expressed cells. Lane M, protein marker. ( B ) The silver-stained protein band indicated by the red arrow was subjected to LC-MS/MS. ( C ) HEK-293T cells were co-transfected with the plasmids encoding PEDV S-Flag and HSC70-Myc, with Flag-tagged or Myc-tagged empty vector as control for 36 h. The supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads or anti-Myc magnetic beads. The precipitated proteins were analyzed by IB. ( D ) Vero cells were co-transfected with the plasmids encoding PEDV S-Flag and HSC70-Myc for 24 h. The cells were visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Pearson’s correlation coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. ( E ) Analyses of KFERQ-like motifs in PEDV S proteins. The KFERQ-like motif typically consists of constant glutamine (Q) flanked on one side of the pentapeptide sequence, followed by one or two positively charged amino residues (lysine, [K] or arginine, [R]), one or two hydrophobic amino residues (phenylalanine, [F]; isoleucine, [I]; leucine, [L]; or valine, [V]), and one negatively charged amino residue (aspartate, [D] and glutamic acid, [E]) . ( F ) HEK-293T cells were co-transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S Q1082A , S Q1117A , S Q1082A/Q1117A , or Flag-tagged empty vector. At 36 h post-transfection, the supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads. The precipitated proteins were analyzed by IB.

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: The KFERQ-like motif (QVDRL) in PEDV S protein is recognized and bound by HSC70. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-ΔCRR or Flag-tagged empty vector for 36 h. The proteins were immunoprecipitated in WCL using anti-Flag magnetic beads, and then the associated proteins were separated by 7.5% SDS-PAGE and stained with silver. The black arrow indicated the expressed S-ΔCRR, and the red arrow indicated a different immunoprecipitated protein band in the S-ΔCRR-expressed cells. Lane M, protein marker. ( B ) The silver-stained protein band indicated by the red arrow was subjected to LC-MS/MS. ( C ) HEK-293T cells were co-transfected with the plasmids encoding PEDV S-Flag and HSC70-Myc, with Flag-tagged or Myc-tagged empty vector as control for 36 h. The supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads or anti-Myc magnetic beads. The precipitated proteins were analyzed by IB. ( D ) Vero cells were co-transfected with the plasmids encoding PEDV S-Flag and HSC70-Myc for 24 h. The cells were visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Pearson’s correlation coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. ( E ) Analyses of KFERQ-like motifs in PEDV S proteins. The KFERQ-like motif typically consists of constant glutamine (Q) flanked on one side of the pentapeptide sequence, followed by one or two positively charged amino residues (lysine, [K] or arginine, [R]), one or two hydrophobic amino residues (phenylalanine, [F]; isoleucine, [I]; leucine, [L]; or valine, [V]), and one negatively charged amino residue (aspartate, [D] and glutamic acid, [E]) . ( F ) HEK-293T cells were co-transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S Q1082A , S Q1117A , S Q1082A/Q1117A , or Flag-tagged empty vector. At 36 h post-transfection, the supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads. The precipitated proteins were analyzed by IB.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Magnetic Beads, SDS Page, Staining, Marker, Liquid Chromatography with Mass Spectroscopy, Control, Software, Sequencing, Residue

Palmitoylation of PEDV S protein prevents its QVDRL motif from being recognized by HSC70. ( A ) HEK-293T cells were transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed cells were treated with or without 40 µM 2-BP. After 36 h post-transfection, the supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads, and the precipitated proteins were analyzed by IB. ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. At 6 h post-transfection, the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP. At 24 h post-transfection, S-Flag, S-ΔCRR, or HSC70 was visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Pearson’s correlation coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. Statistical analysis was carried out using Student’s t -test. ** P < 0.01. ( C ) HEK-293T cells were co-transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S-ΔCRR, S Q1082A , S-ΔCRR Q1082A , or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed and the S Q1082A -overexpressed cells were treated with or without 40 µM 2-BP. Subsequent assays were performed as described for panel A. (D through F) Vero cells were transfected with si- HSC70 or si-NC. At 12 h post-transfection, the cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector, and treated with or without 20 µM 2-BP for another 24 h. The cells were lysed and analyzed by IB. The mean gray values of S protein and HSC70-Myc were quantified using ImageJ software.

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: Palmitoylation of PEDV S protein prevents its QVDRL motif from being recognized by HSC70. ( A ) HEK-293T cells were transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed cells were treated with or without 40 µM 2-BP. After 36 h post-transfection, the supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads, and the precipitated proteins were analyzed by IB. ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. At 6 h post-transfection, the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP. At 24 h post-transfection, S-Flag, S-ΔCRR, or HSC70 was visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Pearson’s correlation coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. Statistical analysis was carried out using Student’s t -test. ** P < 0.01. ( C ) HEK-293T cells were co-transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S-ΔCRR, S Q1082A , S-ΔCRR Q1082A , or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed and the S Q1082A -overexpressed cells were treated with or without 40 µM 2-BP. Subsequent assays were performed as described for panel A. (D through F) Vero cells were transfected with si- HSC70 or si-NC. At 12 h post-transfection, the cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector, and treated with or without 20 µM 2-BP for another 24 h. The cells were lysed and analyzed by IB. The mean gray values of S protein and HSC70-Myc were quantified using ImageJ software.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Magnetic Beads, Staining, Software

Palmitoylation of PEDV S protein prevents its degradation via CMA. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. At 6 h post-transfection, the S-Flag-overexpressed cells were treated with or without 20 µM 2-BP and then treated with NH 4 Cl (0.5 mM). At 24 h post-transfection, S-Flag, S-ΔCRR, or LAMP2A was visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Manders’ overlap coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. Statistical analysis was carried out using Student’s t -test. ** P < 0.01; *** P < 0.001. (B through D) Vero cells were transfected with si- LAMP2A or si-NC. At 12 h post-transfection, the cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector and treated with or without 20 µM 2-BP for another 24 h. The cells were lysed and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: Palmitoylation of PEDV S protein prevents its degradation via CMA. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. At 6 h post-transfection, the S-Flag-overexpressed cells were treated with or without 20 µM 2-BP and then treated with NH 4 Cl (0.5 mM). At 24 h post-transfection, S-Flag, S-ΔCRR, or LAMP2A was visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Manders’ overlap coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. Statistical analysis was carried out using Student’s t -test. ** P < 0.01; *** P < 0.001. (B through D) Vero cells were transfected with si- LAMP2A or si-NC. At 12 h post-transfection, the cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector and treated with or without 20 µM 2-BP for another 24 h. The cells were lysed and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Staining, Software