Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 expression is downregulated in U251 and U87 cells after MUC21 shRNA transfection. (A) Reverse transcription-quantitative PCR assays were performed to measure MUC21 mRNA levels in U251 and U87 cells following control or MUC21 shRNA transfection. (B) Western blot analysis was performed to determine the protein expression of MUC21 in U251 and U87 cells following control or MUC21 shRNA transfection. * P<0.05 vs. shRNA. MUC21, mucin 21; NC, negative control; shRNA, short hairpin RNA.

Article Snippet: MUC21 short hairpin RNA (shRNA; sequence: 5'-GGGTCAGCATAGTCACCAACT-3' and 5'-GCGCTCTGACATGCAGAA-3') and negative control shRNA (shNC; sequence: 5'-TTCTCCGAACGTGTCACGT-3') plasmids were purchased from Santa Cruz Biotechnology, Inc. and transfected into both U251 and U87 cells using Lipofectamine ® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.). shNC was used as a control to compare with MUC21 shRNA (1 μg) transfection. pcDNA3.1-MUC21 was used for overexpression assays and the pcDNA3.1-vector (Addgene, Inc.) was used as control.

Techniques: Expressing, shRNA, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Western Blot, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 promotes GBM cell viability and migration in vitro . (A) Cell viability was assessed after MUC21 depletion in U87 and U251 through colony formation assays. (B) MTT assays were performed to detect the viability of cells transfected with the indicated shRNA. (C) Wound closure assay of GBM cells transfected with the indicated shRNA were performed for the quantification of migrated cells after 24 h. Magnification, x200. Western blotting assays were performed to analyze the expression of (D) Ki-67 and PCNA, and (E) MMP2 and MMP9 in GBM cells transfected with the indicated shRNA. Relative expression was analyzed. * P<0.05 and ** P<0.01 vs. the shNC group. GBM glioblastoma; MUC21, mucin 21; PCNA, proliferating cell nuclear antigen; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: MUC21 short hairpin RNA (shRNA; sequence: 5'-GGGTCAGCATAGTCACCAACT-3' and 5'-GCGCTCTGACATGCAGAA-3') and negative control shRNA (shNC; sequence: 5'-TTCTCCGAACGTGTCACGT-3') plasmids were purchased from Santa Cruz Biotechnology, Inc. and transfected into both U251 and U87 cells using Lipofectamine ® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.). shNC was used as a control to compare with MUC21 shRNA (1 μg) transfection. pcDNA3.1-MUC21 was used for overexpression assays and the pcDNA3.1-vector (Addgene, Inc.) was used as control.

Techniques: Migration, In Vitro, Transfection, shRNA, Wound Closure Assay, Western Blot, Expressing, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 contributes to GBM progression via the STAT3 and AKT signaling pathway. (A) The protein levels of p-STAT3, STAT3, p-AKT and AKT in shMUC21-transfected or control U251 cells were detected by immunoblot assays. (B) The protein levels of MUC21, p-STAT3, STAT3, p-AKT and AKT in MUC21-overexpressing or control U251 cells were detected by immunoblot assays. The relative expression was analyzed. * P<0.05 and ** P<0.01 vs. shNC or Con. Con, control; MUC21, mucin 21; NC, negative control; p-, phosphorylated; shRNA, short hairpin RNA.

Article Snippet: MUC21 short hairpin RNA (shRNA; sequence: 5'-GGGTCAGCATAGTCACCAACT-3' and 5'-GCGCTCTGACATGCAGAA-3') and negative control shRNA (shNC; sequence: 5'-TTCTCCGAACGTGTCACGT-3') plasmids were purchased from Santa Cruz Biotechnology, Inc. and transfected into both U251 and U87 cells using Lipofectamine ® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.). shNC was used as a control to compare with MUC21 shRNA (1 μg) transfection. pcDNA3.1-MUC21 was used for overexpression assays and the pcDNA3.1-vector (Addgene, Inc.) was used as control.

Techniques: Transfection, Control, Western Blot, Expressing, Negative Control, shRNA

Journal: Experimental and Therapeutic Medicine

Article Title: MUC21 induces the viability and migration of glioblastoma via the STAT3/AKT pathway

doi: 10.3892/etm.2022.11260

Figure Lengend Snippet: MUC21 stimulates glioblastoma progression through the STAT3 and AKT pathway in vivo . (A) U251 cells stably transfected with control or MUC21 shRNA vectors were subcutaneously implanted into nude mice. Tumor volume was monitored every week. Isolated tumors were displayed in (A). Tumor growth curves were compared between control and MUC21 depletion groups. Murine weight was also measured. (B) An immunohistochemistry assay was conducted to assess MUC21 protein levels in control or MUC21-depleted groups (scale bar, 200 µm). (C) Western blotting assays were performed to assess the expression levels of the indicated proteins in control or MUC21-depleted tumor tissues isolated from nude mice. * P<0.05 and ** P<0.01 vs. shNC. MUC21, mucin 21; NC, negative control; PCNA, proliferating cell nuclear antigen; p-, phosphorylated; shRNA, short hairpin RNA.

Article Snippet: MUC21 short hairpin RNA (shRNA; sequence: 5'-GGGTCAGCATAGTCACCAACT-3' and 5'-GCGCTCTGACATGCAGAA-3') and negative control shRNA (shNC; sequence: 5'-TTCTCCGAACGTGTCACGT-3') plasmids were purchased from Santa Cruz Biotechnology, Inc. and transfected into both U251 and U87 cells using Lipofectamine ® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.). shNC was used as a control to compare with MUC21 shRNA (1 μg) transfection. pcDNA3.1-MUC21 was used for overexpression assays and the pcDNA3.1-vector (Addgene, Inc.) was used as control.

Techniques: In Vivo, Stable Transfection, Transfection, Control, shRNA, Isolation, Immunohistochemistry, Western Blot, Expressing, Negative Control

Journal: The EMBO Journal

Article Title: Dimer‐specific immunoprecipitation of active caspase‐2 identifies TRAF proteins as novel activators

doi: 10.15252/embj.201797072

Figure Lengend Snippet: A Workflow for mass spectrometry analysis. Casp2pro BiFC cells were treated with mock (DMSO), 20 μM cisplatin, 50 μM etoposide, or 100 nM paclitaxel in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h followed by GFP‐Trap immunoprecipitation. Note the control: Casp2pro BiFC expression was prevented with 1 μg/ml doxycycline, and unconjugated BiFC constructs (VN and VC) were co‐transfected into cells before treatment, similar to experimental samples. Control lysates were then pooled before GFP‐Trap IP. GFP‐Trap beads were subjected to on‐bead protein digestion and analyzed by mass spectrometry. B, C Casp2pro BiFC cells were transfected with siRNA targeting PIDD for 48 h, and knockdown efficiency was confirmed by IB (B). PIDD siRNA transfected cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h. Caspase‐2 BiFC was assessed by flow cytometry. n = 3 independent experiments (means + s.e.m.) (C). D, E Casp2pro BiFC cells were transfected with siRNA targeting RAIDD for 48 h, and knockdown efficiency was confirmed by IB (D). RAIDD siRNA transfected cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h. Caspase‐2 BiFC was assessed by flow cytometry. n = 3 independent experiments (means + s.e.m.) (E). F HeLa cells were transfected with RAIDD siRNA for 48 h and then treated with 20 μM cisplatin for 24 h. Apoptosis was assessed by annexin V staining and flow cytometry. n = 3 independent experiments (means + s.e.m.). G HeLa cells were transfected with Myc‐TRAF2 construct and cultured 48 h for expression. Cell lysates were prepared, followed by Myc‐Trap immunoprecipitation and IB for indicated caspases. H HeLa cells were transfected with caspase‐2(C320A) construct and cultured 48 h for expression. Cell lysates were prepared, followed by IB. I HeLa cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh (left) or treated with 20 ng/ml TNFα (right) for indicated periods. Cell lysates were prepared and analyzed by IB. J Casp2pro BiFC cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh with or without indicated NF‐κB pathway inhibitors, TPCA‐1 or IKK‐16, at indicated concentrations for 24 h. Caspase‐2 BiFC was assessed by flow cytometry. n = 3 independent experiments (means + s.e.m.). Source data are available online for this figure.

Article Snippet: Scramble shRNA (shNT) was a gift from David Sabatini (Addgene plasmid # 1864) (Sarbassov et al , ). shTRAF2 #1 (TRCN0000004573), shTRAF2 #2 (TRCN0000004571), shCasp2 #1 (TRCN0000003508), and shCasp2 #2 (TRCN0000003509) were obtained from Duke Functional Genomics Shared Resource (originally from Sigma). pLKO.1 plasmid and plasmids encoding gag, pol, tat, or VSV‐G were transfected into HEK293T cells with polyethylenimine.

Techniques: Mass Spectrometry, Immunoprecipitation, Control, Expressing, Construct, Transfection, Knockdown, Flow Cytometry, Staining, Cell Culture

Journal: The EMBO Journal

Article Title: Dimer‐specific immunoprecipitation of active caspase‐2 identifies TRAF proteins as novel activators

doi: 10.15252/embj.201797072

Figure Lengend Snippet: A Representative confocal images. Casp2pro BiFC cells were transfected with control siRNA (siCtrl) or siRNA targeting TRAF2 (siTRAF2) for 48 h and then treated with 20 μM cisplatin for 24 h in the presence of 10 μM Q‐VD(OMe)‐OPh. Scale bar indicates 50 μm. B Casp2pro BiFC cells were transfected with the indicated siRNA for 48 h and then treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h. BiFC signal was detected by flow cytometry. n = 3 independent experiments (means + s.e.m.). * P < 0.05 by unpaired two‐tailed t ‐test. C shNT (non‐targeting) or shTRAF2 HeLa cells were treated with 20 μM cisplatin for 24 h, followed by annexin V staining and flow cytometry analysis. n = 3 independent experiments (means + s.e.m.). * P < 0.005 by unpaired two‐tailed t ‐test. D shNT or shCasp2 HeLa cells were analyzed as in (C). n = 3 independent experiments (means + s.e.m.). * P < 0.001 by unpaired two‐tailed t ‐test. E HeLa cells were transfected with siRNA targeting TRAF3 for 48 h, followed by 20 μM cisplatin treatment for 24 h, then stained with annexin V and analyzed by flow cytometry. n = 4 independent experiments (means + s.e.m.). * P < 0.05 by unpaired two‐tailed t ‐test. (A–E, right panel) Knockdown efficiency was assessed by IB. F–I shNT or shTRAF2 #1 HeLa cells were treated with 20 μM cisplatin for 24 h (F and G) or 18 h (H and I), followed by IB for caspase‐2 or caspase‐9 (F), caspase‐3 (G), or BID (H). Active Bax was immunoprecipitated by active Bax‐specific antibody (6A7) and analyzed by IB with total anti‐Bax antibody (I). Source data are available online for this figure.

Article Snippet: Scramble shRNA (shNT) was a gift from David Sabatini (Addgene plasmid # 1864) (Sarbassov et al , ). shTRAF2 #1 (TRCN0000004573), shTRAF2 #2 (TRCN0000004571), shCasp2 #1 (TRCN0000003508), and shCasp2 #2 (TRCN0000003509) were obtained from Duke Functional Genomics Shared Resource (originally from Sigma). pLKO.1 plasmid and plasmids encoding gag, pol, tat, or VSV‐G were transfected into HEK293T cells with polyethylenimine.

Techniques: Transfection, Control, Flow Cytometry, Two Tailed Test, Staining, Knockdown, Immunoprecipitation

Journal: The EMBO Journal

Article Title: Dimer‐specific immunoprecipitation of active caspase‐2 identifies TRAF proteins as novel activators

doi: 10.15252/embj.201797072

Figure Lengend Snippet: Casp2pro BiFC cells were transfected with TRAF2 siRNA for 48 h and then treated with 20 μM cisplatin, 50 μM etoposide, or 100 nM paclitaxel in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h. Caspase‐2 dimerization was assessed by flow cytometry. n = 3 independent experiments (means + s.e.m.). Caspase‐2 or TRAF2 was knocked down by siRNA in BT474 breast cancer cells for 48 h, followed by treatment with 40 μM cisplatin, 100 μM etoposide, or 100 nM paclitaxel for 72 h. Apoptosis was assessed by annexin V staining and flow cytometry. n = 4 independent experiments (means + s.e.m.). shNT or shTRAF2 #1 HeLa cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h. Cells were collected and incubated with BMH for protein crosslinking, and then, lysates were prepared and oligomerized caspase‐2 was detected by IB. Relative signal intensity of oligomerized caspase‐2 was quantified and indicated below each lane (signal was normalized to mock treatment of shNT). TRAF2 CRISPR‐KO HeLa cells were transfected with 50 ng pair of Casp2pro BiFC constructs and cultured for 24 h. Cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h, and caspase‐2 BiFC was assessed by flow cytometry. n = 4 independent experiments (means + s.e.m.). TRAF2 CRISPR‐KO HeLa cells were treated with 20 μM cisplatin for 24 h. Apoptosis was assessed by annexin V staining and flow cytometry. n = 3 independent experiments (means + s.e.m.). Lysates of TRAF2 CRISPR‐KO HeLa (left) or shTRAF2 #1 HeLa cells (right) were prepared, followed by IB. Relative band intensity of TRAF3 was quantified and indicated below each lane [signal was normalized to control (left) or shNT (right)]. Lysates of several clones of TRAF2 CRISPR‐KO HeLa cells were prepared, followed by IB. Control or TRAF2 CRISPR‐KO HeLa cells were transfected with TRAF3 siRNA, followed by Casp2pro BiFC construct transfection. Cells were treated with 20 μM cisplatin in the presence of 10 μM Q‐VD(OMe)‐OPh for 24 h. Caspase‐2 BiFC was assessed by flow cytometry. n = 4 independent experiments (means + s.e.m.). * P < 0.01 (TRAF2 KO #1), P < 0.05 (TRAF2 KO #2) by unpaired two‐tailed t ‐test. n.s.; not significant. Source data are available online for this figure.

Article Snippet: Scramble shRNA (shNT) was a gift from David Sabatini (Addgene plasmid # 1864) (Sarbassov et al , ). shTRAF2 #1 (TRCN0000004573), shTRAF2 #2 (TRCN0000004571), shCasp2 #1 (TRCN0000003508), and shCasp2 #2 (TRCN0000003509) were obtained from Duke Functional Genomics Shared Resource (originally from Sigma). pLKO.1 plasmid and plasmids encoding gag, pol, tat, or VSV‐G were transfected into HEK293T cells with polyethylenimine.

Techniques: Transfection, Flow Cytometry, Staining, Incubation, CRISPR, Construct, Cell Culture, Control, Clone Assay, Two Tailed Test

Journal: The EMBO Journal

Article Title: Dimer‐specific immunoprecipitation of active caspase‐2 identifies TRAF proteins as novel activators

doi: 10.15252/embj.201797072

Figure Lengend Snippet: HEK293T cells were co‐transfected with indicated caspase‐2(C320A)‐Flag (wild type or TIM mutant) and HA‐RAIDD constructs. After 48‐h culture for expression, lysates were prepared, followed by anti‐Flag IP and IB. HEK293T cells were transfected with siRNA targeting caspase‐2 3′‐UTR to deplete endogenous caspase‐2. 24 h post‐siRNA transfection, cells were then transfected with indicated Casp2‐mVenus constructs (full‐length C320A, prodomain, or Δ1–169 C320A (Δprodomain)). Casp2‐mVenus variants were immunoprecipitated by GFP‐Trap, followed by IB for co‐immunoprecipitated TRAF2. Source data are available online for this figure.

Article Snippet: Scramble shRNA (shNT) was a gift from David Sabatini (Addgene plasmid # 1864) (Sarbassov et al , ). shTRAF2 #1 (TRCN0000004573), shTRAF2 #2 (TRCN0000004571), shCasp2 #1 (TRCN0000003508), and shCasp2 #2 (TRCN0000003509) were obtained from Duke Functional Genomics Shared Resource (originally from Sigma). pLKO.1 plasmid and plasmids encoding gag, pol, tat, or VSV‐G were transfected into HEK293T cells with polyethylenimine.

Techniques: Transfection, Mutagenesis, Construct, Expressing, Immunoprecipitation

Journal: The EMBO Journal

Article Title: Dimer‐specific immunoprecipitation of active caspase‐2 identifies TRAF proteins as novel activators

doi: 10.15252/embj.201797072

Figure Lengend Snippet: A Casp2pro BiFC cells were treated with 20 μM cisplatin for 24 h in the presence of 10 μM Q‐VD(OMe)‐OPh, followed by GFP‐Trap IP and IB with anti‐ubiquitin or anti‐GFP antibody. B HeLa cells were treated with 20 μM cisplatin for 24 h in the presence of 10 μM Q‐VD(OMe)‐OPh. Lysates were denatured/renatured and immunoprecipitated with anti‐caspase‐2 antibody or control IgG, followed by IB with anti‐ubiquitin or anti‐caspase‐2 antibody. C HeLa cells were transfected with TRAF2 siRNA for 24 h, then transfected with Casp2pro‐mVenus for 48 h, followed by GFP‐Trap IP and IB. D Casp2(C320A)‐mVenus was co‐expressed with the indicated TRAF2 constructs and then pulled down with GFP‐Trap and analyzed by IB. E In vitro ubiquitylation of recombinant Casp2‐Flag by Myc‐TRAF2 (wild type or ΔRING) purified from HEK293T cells. F, G Casp2pro‐mVenus wild type and indicated lysine mutants were expressed for 24 h in HEK293T cells, followed by GFP‐Trap IP and IB. H HEK293T cells were transfected with Casp2(C320A)‐mVenus (wild type or K15/152/153R (3KR) mutant) constructs for 48 h, followed by GFP‐Trap IP and IB. I HeLa cells were transfected with Casp2pro‐mVenus for 48 h and lysed. Recombinant MBP‐TRAF2 or MBP control proteins were incubated in the lysate for 1 h, followed by amylose pulldown and IB to detect caspase‐2 binding. J In vitro ubiquitylation was performed as in (E), with recombinant Casp2‐Myc protein and Flag‐TRAF2 (wild type or ΔRING) purified from HEK293T cells. After 3‐h incubation at 37°C (Ub reaction (+)) or on ice (No Ub reaction), the reaction was incubated with anti‐Flag beads. Immunoprecipitated and unbound fractions were analyzed by IB. Source data are available online for this figure.

Article Snippet: Scramble shRNA (shNT) was a gift from David Sabatini (Addgene plasmid # 1864) (Sarbassov et al , ). shTRAF2 #1 (TRCN0000004573), shTRAF2 #2 (TRCN0000004571), shCasp2 #1 (TRCN0000003508), and shCasp2 #2 (TRCN0000003509) were obtained from Duke Functional Genomics Shared Resource (originally from Sigma). pLKO.1 plasmid and plasmids encoding gag, pol, tat, or VSV‐G were transfected into HEK293T cells with polyethylenimine.

Techniques: Ubiquitin Proteomics, Immunoprecipitation, Control, Transfection, Construct, In Vitro, Recombinant, Purification, Mutagenesis, Incubation, Binding Assay

Journal: The EMBO Journal

Article Title: Dimer‐specific immunoprecipitation of active caspase‐2 identifies TRAF proteins as novel activators

doi: 10.15252/embj.201797072

Figure Lengend Snippet: shCasp2 #1 cells were transfected with indicated caspase‐2 construct (wild type or 3KR mutant) containing shCasp2‐resistant sequence. Cells were treated with 10 μM cisplatin for 24 h, followed by IB. Relative band intensity of cleaved caspase‐2 was quantified and indicated below each lane (signal was normalized to cisplatin treatment of WT) (top). Transfected cells were treated with 5 μM cisplatin for 24 h, and then analyzed by annexin V staining and flow cytometry. n = 4 independent experiments (means + s.e.m.) (bottom). HeLa cells were transfected with indicated Myc‐TRAF2 construct (wild type or RING domain mutant) for 48 h. Apoptosis was analyzed by annexin V staining and flow cytometry. n = 3 independent experiments (means + s.e.m.) (left). Expression of Myc‐TRAF2 was confirmed by IB (right). HeLa shNT, shCasp2 #1, or shCasp2 #2 cells were transfected as in (B), and apoptosis was analyzed by annexin V staining and flow cytometry. n = 3 independent experiments (means + s.e.m.). Source data are available online for this figure.

Article Snippet: Scramble shRNA (shNT) was a gift from David Sabatini (Addgene plasmid # 1864) (Sarbassov et al , ). shTRAF2 #1 (TRCN0000004573), shTRAF2 #2 (TRCN0000004571), shCasp2 #1 (TRCN0000003508), and shCasp2 #2 (TRCN0000003509) were obtained from Duke Functional Genomics Shared Resource (originally from Sigma). pLKO.1 plasmid and plasmids encoding gag, pol, tat, or VSV‐G were transfected into HEK293T cells with polyethylenimine.

Techniques: Transfection, Construct, Mutagenesis, Sequencing, Staining, Flow Cytometry, Expressing

Journal: Frontiers in Immunology

Article Title: Prognostic stratification of sepsis through DNA damage response based RiskScore system: insights from single-cell RNA-sequencing and transcriptomic profiling

doi: 10.3389/fimmu.2024.1345321

Figure Lengend Snippet: Validation of characteristic genes in vivo (A) Relative expression levels of DDR-related genes in control and sepsis groups (n=4 in each group). (B) Relative expression levels of ARL4C in control, sepsis, sepsis+Ad-shNC, and sepsis+Ad-shARL4C group (n=8 in each group). (C) Expression levels of inflammatory factors (IL-1β, TNF-α, IL-10, and IL-18) in the peripheral blood of rat with sepsis+Ad-shNC and sepsis+Ad-shARL4C (n=8 in each group). (D) Survival status of rats in each group (n=10 in each group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. NS, no significant difference.

Article Snippet: In summary, adenoviral vectors containing shRNA sequences specifically targeting the ARL4C gene in rats (Ad-shARL4C) alongside a non-targeting control shRNA sequence (Ad-shNC), both sourced from RiboBio in Guangzhou, China, were utilized.

Techniques: Biomarker Discovery, In Vivo, Expressing, Control

Journal: Frontiers in Immunology

Article Title: Prognostic stratification of sepsis through DNA damage response based RiskScore system: insights from single-cell RNA-sequencing and transcriptomic profiling

doi: 10.3389/fimmu.2024.1345321

Figure Lengend Snippet: Validation of ARL4C in vitro . (A) Flow cytometry detected the apoptosis rate in the Control, Sepsis, Sepsis+Lv-shNC, and Sepsis+Lv-shARL4C groups. (n=4 in each group) (B) Flow cytometry detected the ROS production in the Control, Sepsis, Sepsis+Lv-shNC, and Sepsis+Lv-shARL4C groups. (n=4 in each group), *** p < 0.001, **** p < 0.001. NS, no significant difference.

Article Snippet: In summary, adenoviral vectors containing shRNA sequences specifically targeting the ARL4C gene in rats (Ad-shARL4C) alongside a non-targeting control shRNA sequence (Ad-shNC), both sourced from RiboBio in Guangzhou, China, were utilized.

Techniques: Biomarker Discovery, In Vitro, Flow Cytometry, Control