A-21277

antibody from Invitrogen Antibodies

Targeting: ELAVL1 Hua, HUR, MelG

Western blot

Antibody data

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Validation data

ELAVL1 protein structure - A-21277 shown in red.

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 CSE, HRV, or HRV+CSE do not alter AUF-1, KHSRP and HuR expression in HBE cells. HBE cells were treated with medium (control), CSE, HRV or the combination for the times indicated. Total cell lysates were harvested and analyzed via immunoblotting. Membranes were probed with specific AUF-1 (A) , KHSRP (B) or HuR (C) antibodies, then were subsequently stripped and re-probed with an antibody to GAPDH to ensure equal protein loading. Data are representative of 3 separate experiments. Relative expression levels of each protein of interest from the 3 experiments were assessed by densitometry.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Effects of HuR-targeting siRNA on CXCL8 protein expression in HBE cells. HBE cells were treated for 24 h prior to harvesting cell supernatants for analysis with ELISA and whole cell lysates for analysis with immunoblotting. To determine the level of HuR protein knock-down membranes were probed with a specific HuR antibody, then were subsequently stripped and re-probed with an antibody to GAPDH to ensure equal protein loading (A ; representative of 3 separate experiments ) . Densitometry analysis with % knock-down relative to HRV+CSE alone is also shown (A ; n=3 ) . Cell supernatants were analyzed for CXCL8 protein (B ; n=6 ) . Data are presented as mean +- SEM. Lipid denotes transfection reagent alone. NT denotes non-targeting control siRNA. Asterisks denote a significant difference between the specified treatments (***p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 HuR knockdown affects HRV+CSE-induced, but not basal, CSE or HRV-induced CXCL8 protein expression in HBE cells. HBE cells were treated for 24 h prior to harvesting cell supernatants for analysis with ELISA and whole cell lysates for analysis with immunoblotting. To determine the level of HuR protein knock-down membranes were probed with a specific HuR antibody, then were subsequently stripped and re-probed with an antibody to GAPDH to ensure equal protein loading (A ; representative of 3 separate experiments ) . Cell supernatants were analyzed for CXCL8 protein (B ; n=3 ) . Data are presented as mean +- SEM. Lipid denotes transfection reagent alone. NT denotes non-targeting control siRNA. Asterisks denote a significant difference between the specified treatments (*p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Knockdown of IGF2BPs decreases mRNA stability ( a ) Cumulative distribution of mRNA half-lives of non-target or IGF2BP high-confidence target genes in HepG2 (left) and Hela (right) cells. ( b ) Distribution of mRNA half-lives in IGF2BP3 high-confidence targets in HepG2 cells with shIGF2BP3 or shNS. ( c ) Cumulative distribution of mRNA half-life of IGF2BP3 high-confidence targets in shIGF2BP3 or shNS HepG2 cells. mRNA half-life analyses in a, b and c were repeated twice. ( d and e ) Reducing MYC mRNA half-life by silencing IGF2BPs ( d ) or m 6 A writers ( e ) in HepG2 cells. Values are mean+-s.d. of n =3 independent experiments. ( f ) Co-Immunoprecipitation and Western blotting showing the binding of mRNA stabilizers with FLAG-tagged IGF2BPs in HEK293T cells, representative of 3 independent experiments. ( g and h ) Co-localization of IGF2BP proteins with HuR ( g ) or DCP1A ( h ) in Hela cells. Arrows indicate co-localization in cytoplasmic granules. Scale bar=10mum. Images were representative of 3 independent experiments. P values were calculated using two-sided Wilcoxon and Mann-Whitney test in a , b , c . Unprocessed scans of western blot analysis are available in Supplementary Figure 8 . Source data of d and e are in Supplementary Table 3 .