- PA5-17281 - Invitrogen Antibodies EGFR antibody

Submitted references METTL14 promotes tumorigenesis by regulating lncRNA OIP5-AS1/miR-98/ADAMTS8 signaling in papillary thyroid cancer.

Zhang X, Li D, Jia C, Cai H, Lv Z, Wu B
Cell death & disease 2021 Jun 15;12(6):617

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of EGFR was performed using 90% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-EGFR(Tyr 1086) Polyclonal Antibody (Product # PA5-17281) at the dilution of 1:100 in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membranous localization. Panel e represents cells treated with antagonist, Afatinib (1uM for 6hrs) followed by EGF (200 ng/mL for 10 minutes), showing no signal. Panel f shows untreated cells with no signal. Panel g represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 OIP5-AS1 and ADAMTS8 regulate PTC cell proliferation, migration/invasion and activate EGFR and MEK/ERK pathways in vitro. A , B OIP5-AS1 and ADAMTS8 expression in four PTC cell lines (TPC-1, BHP5-16, K1, and BCPAP), and in a normal human thyroid epithelial cell line Nthy-ori3-1 were analyzed by qRT-PCR ( n = 3). C , D qRT-PCR analysis of ADAMTS8 expression levels in K1 cells and TPC-1 cells, K1 cells were transfected with si-NC/si-OIP5-AS1, while TPC-1 cells were transfected with pcDNA-3.1/OIP5-AS1 ( n = 3). E-I K1 cells were co-transfected with OIP5-AS1 and si-ADAMTS8, while TPC-1 cells were co-transfected with si-OIP5-AS1/ADAMTS8 ( n = 3). E , F MTT and colony-forming growth assays were performed to determine the proliferation of K1 and TPC-1 cells harboring the different vectors indicated ( n = 3). G Transwell assays were performed to determine the migration and invasion capacity of K1 and TPC-1 cells ( n = 3). Scale bar = 50 mum. H Wound healing assays were performed to assess the migratory capacity of K1 and TPC-1 cells ( n = 3). I Migration and invasion abilities (fold change of migrated or invaded) were calculated, compared with the different vectors in K1 and TPC-1cells. The migratory activity (wound healing) was calculated and compared with those at 0 h ( n = 3). J Western blot analysis of phosphorylated EGFR and total EGFR in TPC-1 and K1 cells ( n = 3). K Phosphorylated and total Akt, ERK, and MEK levels measured by western blot analysis in TPC-1 and K1 cell

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 7 METTL14 promotes PTC cell proliferation, migration/invasion, and activate downstream EGFR, Akt, and MEK/ERK pathways through lncRNA OIP5-AS1. A , B MTT and colony-forming growth assays were performed to determine the proliferation ability of K1 and TPC-1 cells ( n = 3). C Transwell assays were performed to determine the migration and invasion capacity of K1 and TPC-1 cells. Scale bars = 50 mum. The migration and invasion abilities (fold change of migrated or invaded) were calculated and compared to the different vectors in K1 and TPC-1 cells ( n = 3). D Wound healing assays were performed to assess the migratory capacity of K1 and TPC-1 cells. The migratory activity (wound healing) was calculated and compared to vectors at 0 h ( n = 3). E Western blot analysis of phosphorylated EGFR and total EGFR in TPC-1 and K1 cells ( n = 3). F Phosphorylated and total Akt, ERK, and MEK levels measured by western blot analysis in TPC-1 and K1 cells ( n = 3). * p < 0.05 and ** p < 0.01.

PA5-17281

Antibody data