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ProSci Incorporated trim21 e 11
Trim21 E 11, supplied by ProSci Incorporated, 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/trim21+e+11/TRIM21+Antibody/pm33937329-40-15-32
Average 93 stars, based on 1 article reviews

trim21 e 11 - by Bioz Stars, 2026-09

93/100 stars

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Article Title: TRIM21, a New Component of the TRAIL-Induced Endogenous Necrosome Complex.
Article Snippet: .. Used antibodies were antibodies to caspase-8 clone C15 (ALX-804-429) (Alexis Biochemicals), FADD (556402) (Becton Dickinson), TRIM21 E-11 (sc-48430), Hsc70 (sc7298) (Santa-Cruz), β-actin (A5316) (Sigma), RIPK3 (48- 909), DR4 (1167), and DR5 (2019) (ProSci Incorporated), RIPK1 (D94C12) XP (3493S) (Cell Signaling), phosphoMLKL (ab187091) (Abcam), RIPK3 (PA1-41533) (Pierce), phospho-RIP1 (Ser166) (D1L3S), and phospho-RIP3 (Ser227) (D6W2T) (Cell Signaling) (this rabbit monoclonal antibody to P-RIPK3 recognizes two bands on western blot), FADD (06711) (Millipore), goat polyclonal antibodies to RIPK3 (clone N-14) (sc-47368), TRIM21 M-20 (sc-21367) (Santa-Cruz) and rat monoclonal antibodies to MLKL (MABC604) (Millipore). .. Horseradish-peroxydase-conjugated secondary antibodies were provided by Dako and Clean Blot IP detection kit provided by Life Technologies.

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TRIM21 reverses the decrease in cell proliferation and colony formation caused by UV irradiation. ( A ) MCF7 cells transfected with TRIM21-expressing plasmid construct were exposed to 10 J/m 2 of UVC radiation or not exposed, and allowed to grow for 72 h post treatment. Cells were harvested and MTT assay was performed at indicated time points. ( B ) MCF7 cells exposed to UV were seeded and colonies were counted after 12–14 days by crystal violet staining (Left panel). Normalized CFUs from three independent experiments were plotted (Right panel). ( C ) MCF7 cells transfected with different plasmid constructs expressing TRIM21 and/or HuR were allowed to grow post transfection. MTT assay was performed at indicated time points. ( D ) Transfected MCF7 cells as indicated in C were seeded and colonies were counted after 12–14 days by crystal violet staining (Left panel). Normalized CFUs from three independent experiments were plotted (Right panel). *Signifies a p value ≤ 0.05, **signifies a p value ≤ 0.01, ***signifies a p value ≤ 0.005 (paired two-tailed or one-tailed t-test).

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: TRIM21 reverses the decrease in cell proliferation and colony formation caused by UV irradiation. ( A ) MCF7 cells transfected with TRIM21-expressing plasmid construct were exposed to 10 J/m 2 of UVC radiation or not exposed, and allowed to grow for 72 h post treatment. Cells were harvested and MTT assay was performed at indicated time points. ( B ) MCF7 cells exposed to UV were seeded and colonies were counted after 12–14 days by crystal violet staining (Left panel). Normalized CFUs from three independent experiments were plotted (Right panel). ( C ) MCF7 cells transfected with different plasmid constructs expressing TRIM21 and/or HuR were allowed to grow post transfection. MTT assay was performed at indicated time points. ( D ) Transfected MCF7 cells as indicated in C were seeded and colonies were counted after 12–14 days by crystal violet staining (Left panel). Normalized CFUs from three independent experiments were plotted (Right panel). *Signifies a p value ≤ 0.05, **signifies a p value ≤ 0.01, ***signifies a p value ≤ 0.005 (paired two-tailed or one-tailed t-test).

Article Snippet: This was followed by immunoblotting using anti- HuR (3A2, Santacruz Biotechnology), p53 (DO-1, Santa Cruz Biotechnology), TRIM21 (E-11, Santa Cruz Biotechnology) or (D101D, Cell Signaling Technologies), Myc (71D10, Cell Signaling Technologies), GAPDH (FL-335, Santacruz Biotechnology), and β-Actin (A00730, Genscript) primary antibodies.

Techniques: Irradiation, Transfection, Expressing, Plasmid Preparation, Construct, MTT Assay, Staining, Two Tailed Test, One-tailed Test

Posttranscriptional regulation of TRIM21 by HuR. ( A ) Sequence of 3′UTR of TRIM21 mRNA indicating a putative U-rich HuR binding site. ( B ) mFold structure of 3′UTR of TRIM21 mRNA including HuR binding site (highlighted in yellow). ( C ) Immunoblots of lysates of MCF7 cells transfected with two increasing concentrations of Myc-tagged-HuR (WT) expressing constructs probed with TRIM21, HuR, and β-Actin antibodies (upper panel). qRT-PCR of total RNA from the same cell lysate with TRIM21- and GAPDH-specific primers (lower panel). ( D ) Immunoblots of lysates of MCF7 cells transfected with two increasing concentrations of siRNA against endogenous HuR probed with TRIM21, HuR, and GAPDH antibodies (upper panel). qRT-PCR of total RNA from the same cell lysate with TRIM21- and GAPDH specific primers (lower panel). Mean ± SD from three independent experiments, each with two technical replicates are represented in all graphs.

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: Posttranscriptional regulation of TRIM21 by HuR. ( A ) Sequence of 3′UTR of TRIM21 mRNA indicating a putative U-rich HuR binding site. ( B ) mFold structure of 3′UTR of TRIM21 mRNA including HuR binding site (highlighted in yellow). ( C ) Immunoblots of lysates of MCF7 cells transfected with two increasing concentrations of Myc-tagged-HuR (WT) expressing constructs probed with TRIM21, HuR, and β-Actin antibodies (upper panel). qRT-PCR of total RNA from the same cell lysate with TRIM21- and GAPDH-specific primers (lower panel). ( D ) Immunoblots of lysates of MCF7 cells transfected with two increasing concentrations of siRNA against endogenous HuR probed with TRIM21, HuR, and GAPDH antibodies (upper panel). qRT-PCR of total RNA from the same cell lysate with TRIM21- and GAPDH specific primers (lower panel). Mean ± SD from three independent experiments, each with two technical replicates are represented in all graphs.

Techniques: Sequencing, Binding Assay, Western Blot, Transfection, Expressing, Construct, Quantitative RT-PCR

$HuR enhances the translation of TRIM21 mRNA. ( A ) MCF7 cells transfected with two increasing concentrations of Myc-tagged-HuR (WT) expressing constructs were treated with or without CHX. Immunoblotting was performed from the lysates using TRIM21, Myc-tag, and GAPDH specific antibodies. ( B ) Ribosomal fractions from MCF7 cells, either mock transfected or transfected with Myc-tagged HuR (WT) expressing constructs, were analysed by sucrose density gradient fractionation (upper panels). Total RNAs isolated from the indicated fractions were analysed by semi-quantitative RT-PCR using TRIM21 and GAPDH primers (middle panels). Percentage of band intensity was plotted against the fraction number (lower panels).$

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: HuR enhances the translation of TRIM21 mRNA. ( A ) MCF7 cells transfected with two increasing concentrations of Myc-tagged-HuR (WT) expressing constructs were treated with or without CHX. Immunoblotting was performed from the lysates using TRIM21, Myc-tag, and GAPDH specific antibodies. ( B ) Ribosomal fractions from MCF7 cells, either mock transfected or transfected with Myc-tagged HuR (WT) expressing constructs, were analysed by sucrose density gradient fractionation (upper panels). Total RNAs isolated from the indicated fractions were analysed by semi-quantitative RT-PCR using TRIM21 and GAPDH primers (middle panels). Percentage of band intensity was plotted against the fraction number (lower panels).

Techniques: Transfection, Expressing, Construct, Western Blot, Fractionation, Isolation, Quantitative RT-PCR

HuR interacts with 3′UTR of TRIM21 mRNA in cells and in vitro. ( A ) MCF7 cell lysates were immunoprecipitated using nonspecific and HuR-specific antibodies and immunoblotted using HuR and β-Actin antibodies (left panel). qRT-PCR from the total RNA isolated from the immunoprecipitated samples was performed using TRIM21 and GAPDH specific primers (Right panel). Data represents mean ± SD values from 3 independent experiments. **Signifies a p value ≤ 0.01, (paired two-tailed t-test). ( B ) 32 P-labelled 3′UTR of TRIM21 mRNA was incubated with MCF7 S10 cytoplasmic lysate, UV-crosslinked and RNaseA digested. The RNA–protein complexes were immunoprecipitated using HuR-specific and non-specific antibodies and electrophoresed on 12% SDS-PAGE gel. ( C ) Schematic diagram of the TRIM21 wild-type ( TRIM21 WT) and TRIM21 mutant ( TRIM21 MT) 3′UTR RNAs. ( D ) 32 P-labelled 3′UTR of TRIM21 mRNA was incubated with three increasing concentrations of purified his-tagged HuR, UV-crosslinked and RNaseA digested. The RNA–protein complexes were resolved on 12% SDS-PAGE gel (left panel). Purified HuR was incubated with three increasing concentrations of unlabelled TRIM21 3′UTR prior to the incubation with 32 P-labelled TRIM21 3′UTR. UV-crosslinking was done followed by RNaseA digestion and RNA–protein complexes were resolved using 12% SDS-PAGE gel (Right panel). ( E ) 32 P-labelled TRIM21 WT and MT 3′UTRs were incubated with three increasing concentrations of purified his-tagged HuR, UV-crosslinked and RNaseA digested. The RNA–protein complexes were resolved on 12% SDS-PAGE gel.

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: HuR interacts with 3′UTR of TRIM21 mRNA in cells and in vitro. ( A ) MCF7 cell lysates were immunoprecipitated using nonspecific and HuR-specific antibodies and immunoblotted using HuR and β-Actin antibodies (left panel). qRT-PCR from the total RNA isolated from the immunoprecipitated samples was performed using TRIM21 and GAPDH specific primers (Right panel). Data represents mean ± SD values from 3 independent experiments. **Signifies a p value ≤ 0.01, (paired two-tailed t-test). ( B ) 32 P-labelled 3′UTR of TRIM21 mRNA was incubated with MCF7 S10 cytoplasmic lysate, UV-crosslinked and RNaseA digested. The RNA–protein complexes were immunoprecipitated using HuR-specific and non-specific antibodies and electrophoresed on 12% SDS-PAGE gel. ( C ) Schematic diagram of the TRIM21 wild-type ( TRIM21 WT) and TRIM21 mutant ( TRIM21 MT) 3′UTR RNAs. ( D ) 32 P-labelled 3′UTR of TRIM21 mRNA was incubated with three increasing concentrations of purified his-tagged HuR, UV-crosslinked and RNaseA digested. The RNA–protein complexes were resolved on 12% SDS-PAGE gel (left panel). Purified HuR was incubated with three increasing concentrations of unlabelled TRIM21 3′UTR prior to the incubation with 32 P-labelled TRIM21 3′UTR. UV-crosslinking was done followed by RNaseA digestion and RNA–protein complexes were resolved using 12% SDS-PAGE gel (Right panel). ( E ) 32 P-labelled TRIM21 WT and MT 3′UTRs were incubated with three increasing concentrations of purified his-tagged HuR, UV-crosslinked and RNaseA digested. The RNA–protein complexes were resolved on 12% SDS-PAGE gel.

Techniques: In Vitro, Immunoprecipitation, Quantitative RT-PCR, Isolation, Two Tailed Test, Incubation, SDS Page, Mutagenesis, Purification

Effect of HuR on TRIM21 mRNA translation is mediated through its 3′UTR. ( A ) Schematic representation of different TRIM21 luciferase reporter constructs. ( B ) Luciferase reporter assay was performed with the MCF7 cells transfected with different Firefly luciferase constructs, cotransfected with two increasing concentrations of WT Myc-tagged HuR expressing construct. ( C ) HuR and control IgG immunoprecipitation was performed with lysates of cells transfected with Firefly luciferase (having wild type or mutant TRIM21 mRNA 3′UTR) reporter gene construct and a Renilla luciferase construct (as transfection control). qRT-PCR from the total RNA isolated from the immunoprecipitated samples was performed using F-Luc and R-Luc specific primers. The ratio of F-Luc/R-Luc RNA level was used to calculate fold change of HuR-immunoprecipitated RNA vs IgG-immunoprecipitated RNA in wild-type and mutant TRIM21 3′UTR transfected cells. ( D ) Luciferase reporter assay was performed with the MCF7 cells transfected with different Firefly luciferase constructs, cotransfected with two increasing concentrations of siRNA against endogenous HuR . F-Luc values are normalized to R-Luc values as transfection control. Data represents mean ± SD values from 3 independent experiments. *Signifies a p value ≤ 0.05, **signifies a p value ≤ 0.01 (paired two-tailed t-test).

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: Effect of HuR on TRIM21 mRNA translation is mediated through its 3′UTR. ( A ) Schematic representation of different TRIM21 luciferase reporter constructs. ( B ) Luciferase reporter assay was performed with the MCF7 cells transfected with different Firefly luciferase constructs, cotransfected with two increasing concentrations of WT Myc-tagged HuR expressing construct. ( C ) HuR and control IgG immunoprecipitation was performed with lysates of cells transfected with Firefly luciferase (having wild type or mutant TRIM21 mRNA 3′UTR) reporter gene construct and a Renilla luciferase construct (as transfection control). qRT-PCR from the total RNA isolated from the immunoprecipitated samples was performed using F-Luc and R-Luc specific primers. The ratio of F-Luc/R-Luc RNA level was used to calculate fold change of HuR-immunoprecipitated RNA vs IgG-immunoprecipitated RNA in wild-type and mutant TRIM21 3′UTR transfected cells. ( D ) Luciferase reporter assay was performed with the MCF7 cells transfected with different Firefly luciferase constructs, cotransfected with two increasing concentrations of siRNA against endogenous HuR . F-Luc values are normalized to R-Luc values as transfection control. Data represents mean ± SD values from 3 independent experiments. *Signifies a p value ≤ 0.05, **signifies a p value ≤ 0.01 (paired two-tailed t-test).

Techniques: Luciferase, Construct, Reporter Assay, Transfection, Expressing, Control, Immunoprecipitation, Mutagenesis, Quantitative RT-PCR, Isolation, Two Tailed Test

HuR-mediated translational upregulation of TRIM21 is prevented by DHTS-I. ( A ) Chemical Structure of dihydrotanshinone-I (DHTS-I). ( B ) HuR was RNA-immunoprecipitated from the lysates of the MCF7 cells treated or untreated with DHTS-I. qRT-PCR from the total RNA isolated from the immunoprecipitates was carried out using TRIM21 and GAPDH primers. ( C ) Luciferase reporter assay was performed with the MCF7 cells transfected with different Firefly luciferase constructs, treated with or without DHTS-I. F-Luc values are normalized to R-Luc values as transfection control. Data represents mean ± SD values from 3 independent experiments. *Signifies a p value ≤ 0.05, (paired one-tailed t-test). ( D ) MCF7 cells transfected with Myc-tagged HuR expressing construct, treated with or without DHTS-I were lysed and immunoblotted using TRIM21, p53, HuR and GAPDH antibodies.

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: HuR-mediated translational upregulation of TRIM21 is prevented by DHTS-I. ( A ) Chemical Structure of dihydrotanshinone-I (DHTS-I). ( B ) HuR was RNA-immunoprecipitated from the lysates of the MCF7 cells treated or untreated with DHTS-I. qRT-PCR from the total RNA isolated from the immunoprecipitates was carried out using TRIM21 and GAPDH primers. ( C ) Luciferase reporter assay was performed with the MCF7 cells transfected with different Firefly luciferase constructs, treated with or without DHTS-I. F-Luc values are normalized to R-Luc values as transfection control. Data represents mean ± SD values from 3 independent experiments. *Signifies a p value ≤ 0.05, (paired one-tailed t-test). ( D ) MCF7 cells transfected with Myc-tagged HuR expressing construct, treated with or without DHTS-I were lysed and immunoblotted using TRIM21, p53, HuR and GAPDH antibodies.

Techniques: Immunoprecipitation, Quantitative RT-PCR, Isolation, Luciferase, Reporter Assay, Transfection, Construct, Control, One-tailed Test, Expressing

Model of HuR-mediated translation regulation of pro-oncogenic factor TRIM21 influencing the expression of p53 under genotoxic stress. ( A ) Plot of % of patients with different cancers with high level of TRIM21 expression (Protein Expression Summary from Human Protein Atlas, v19.proteinatlas.org/ENSG00000132109-TRIM21/pathology). ( B ) Proposed model of HuR-mediated translation activation of TRIM21. HuR overexpression enhances the rate of translation of TRIM21 mRNA which can be reversed by DHTS-I treatment. ( C ) Network diagram representing the regulation of p53 mediated by HuR, miR-125b, TRIM21 and Mdm2 in response to UV-induced DNA damage, indicating the negative feedback loop between TRIM21 and HuR.

Journal: Scientific Reports

Article Title: Negative feedback regulation by HuR controls TRIM21 expression and function in response to UV radiation

doi: 10.1038/s41598-020-68646-3

Figure Lengend Snippet: Model of HuR-mediated translation regulation of pro-oncogenic factor TRIM21 influencing the expression of p53 under genotoxic stress. ( A ) Plot of % of patients with different cancers with high level of TRIM21 expression (Protein Expression Summary from Human Protein Atlas, v19.proteinatlas.org/ENSG00000132109-TRIM21/pathology). ( B ) Proposed model of HuR-mediated translation activation of TRIM21. HuR overexpression enhances the rate of translation of TRIM21 mRNA which can be reversed by DHTS-I treatment. ( C ) Network diagram representing the regulation of p53 mediated by HuR, miR-125b, TRIM21 and Mdm2 in response to UV-induced DNA damage, indicating the negative feedback loop between TRIM21 and HuR.

Techniques: Expressing, Activation Assay, Over Expression

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