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Image Search Results
Journal: International Journal of Molecular Sciences
Article Title: A Positive Feedback DNA-PK/MYT1L-CXCR1-ERK1/2 Proliferative Signaling Loop in Glioblastoma
doi: 10.3390/ijms26094398
Figure Lengend Snippet: Transcriptional control of CXCR1 by MYT1L and DNA-PK. ( A ) Whole cellular lysates were prepared from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to Western blot analysis of MYT1L; GAPDH served as a loading control. ( B ) Total RNA was isolated from BJ-5ta, A-172, M059J, M059K, IMR-5, IMR-32, SH-SY5Y, SK-N-AS, SK-N-BE(2), SK-N-MC, and SK-N-SH cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( C ) Total RNA was isolated from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to the qRT-PCR analysis of CXCR1 that was performed in triplicate. ( D , E ) Whole cellular lysates were prepared from HEK293-MYT1L, HEK293-GFP, M059J-MYT1L, M059J-GFP, M059K-MYT1L, and M059K-GFP cells and subjected to Western blot analysis of CXCR1 and CXCR2; actin and GAPDH served as a loading control. Note that the images of blots have been cropped and aligned in the same order as panels “( C )” and “( E )”. ( F ) A diagram of wild-type and mutant CXCR1 promoter/reporter constructs. ( G ) HEK293, M069J, and M059K cells were cotransfected with either an empty vector or pGL3-wtCXCR1-luc or pGL3-mtCXCR1 −2114/−2103 -luc reporter plasmid in combination with pCMV6-MYT1L and pRL-TK; at 24 h after transfection, the luciferase activity was measured in duplicate as described in “Methods”. * indicates p < 0.05.
Article Snippet: HEK293, M059J, and M059K cells grown to 90% confluency in a 6-well plate were transiently cotransfected with 0.5 μg of either
Techniques: Control, Western Blot, Isolation, Quantitative RT-PCR, Mutagenesis, Construct, Plasmid Preparation, Transfection, Luciferase, Activity Assay