Fig. 4: TRMP specifically modulates protein levels of p27. | Cell Death & Disease

Fig. 4: TRMP specifically modulates protein levels of p27.

From: TRMP, a p53-inducible long noncoding RNA, regulates G1/S cell cycle progression by modulating IRES-dependent p27 translation

Fig. 4: TRMP specifically modulates protein levels of p27.The alternative text for this image may have been generated using AI.

a A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. Forty-eight hours after infection, total RNA was subjected to real-time RT-PCR analysis to examine mRNA levels of the indicated genes. Data shown are mean ± SD (n = 3). b A549 cells were infected with lentiviruses expressing either control RNA or TRMP. Forty-eight hours after infection, total RNA was subjected to real-time RT-PCR analysis to examine mRNA levels of the indicated genes. Data shown are mean ± SD (n = 3). c A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. Forty-eight hours after infection, cell lysates were analyzed by western blotting with the indicated antibodies. The band intensities were quantified by using ImageJ software. The ratio of p27 to GAPDH is presented in Supplementary Figure S3C. d A549 cells were infected with lentiviruses expressing either control RNA or TRMP. Forty-eight hours after infection, cell lysates were analyzed by western blotting with anti-p27 antibody. The band intensities were quantified by using ImageJ software. The ratio of p27 to GAPDH is presented in Supplementary Figure S3D. e, f A549 cells were infected with lentiviruses expressing either control RNA or TRMP. e Forty-eight hours after infection, cells were treated with cycloheximide (CHX, 20 μg/ml) for the indicated periods of time. Cell lysates were then analyzed by western blotting. f The band intensities were quantified by using ImageJ software. The ratio of p27 to GAPDH was also presented. g, h A549 cells were infected with lentiviruses expressing control shRNA, TRMP shRNA#1, or TRMP shRNA#2. g Forty-eight hours after infection, cells were treated with cycloheximide (CHX, 20 μg/ml) for the indicated periods of time. Cell lysates were then analyzed by western blotting to determine p27 protein half-life. h The band intensities were quantified by using ImageJ software. The ratio of p27 to GAPDH was also presented. i Lysates from A549 cells were incubated with either sense or antisense biotin-labeled DNA oligomers corresponding to TRMP, followed by the pull-down experiments using streptavidin-coated beads. The pull-downed complexes were subjected to real-time RT-PCR analysis to detect p27 mRNA levels

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