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Fig. 2 | BMC Cancer

Fig. 2

From: PCAF-mediated acetylation of Lin28B increases let-7 biogenesis in lung adenocarcinoma H1299 cells

Fig. 2

Lin28B interacts with PCAF via its cold shock domain. a Co-IP of the co-transfected PCAF-FLAG and Lin28B-GFP expression constructs in HEK293T cells. WCEs were IP-ed with anti-GFP antibody, and immunoblotted (IB) with the antibodies shown on the right. WCEs without immunoprecipitation are shown as Input. b GST-pulldown assay. Lin28B-GFP or GFP alone was individually incubated with WCEs of HEK293T cells ectopically expressing FLAG-PCAF, and the samples were subjected to gel electrophoresis and were blotted with antibodies shown on the right. c Schematic representation of the truncation fragments of Lin28B. Digits indicate the positions of the amino acids at the terminus of each fragment. d Domain-specific binding of Lin28B with PCAF. Each of the expression constructs of Lin28B and its individually truncated fragments was tagged with GFP (labeled as L1~L7) and individually co-transfected with PCAF-FLAG in HEK293T cells. WCEs of each transfected sample was IP-ed with anti-GFP antibody and then subjected to gel electrophoresis and was blotted with antibodies shown on the right. c GST-pull down assay identifying the interactions between specific Lin28B domains and PCAF in vitro. As shown in (d), each of the GST-tagged Lin28B and the related samples indicated at top was incubated with WCEs containing FLAG-tagged PCAF, and samples were then analyzed by western blotting with antibodies against GST and FLAG, as indicated. f Co-localization of Lin28B and PCAF in human cells. Immunofluorescence images showing endogenous Lin28B and PCAF in H1299 cells

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