Fig. 2: FHL2 is not modified by PARP12 in vitro and in vivo. | Cell Death & Disease

Fig. 2: FHL2 is not modified by PARP12 in vitro and in vivo.

From: PARP12 (ARTD12) suppresses hepatocellular carcinoma metastasis through interacting with FHL2 and regulating its stability

Fig. 2

a, b FHL2 is not modified by PARP12 as examined by an in vitro ADP-ribosylation assay. GST-FHL2 (1 µg) was incubated with His-PARP12 (0.5 µg) in the presence or absence of biotin-NAD+ or β-NAD+ (25 µM) at 30 °C for 30 min. The reactions were stopped by the addition of 2 × SDS loading buffer and boiled at 98 °C for 10 min. The samples were analyzed by Western blot using streptavidin-HRP (a) or anti-ADP-ribose (b) antibody. *: non-specific bands. c FHL2 is not mono-ADP-ribosylated in vivo. GFP-PARP12 WT, GFP-PARP12 MT (H564Y G565A), and GFP-FHL2 were transfected into HEK293T cells. Whole-cell lysates were subjected to pull-down assays using the His-tagged macro domains 1–3 of PARP14 coupled with Ni Sepharose. Proteins were separated by SDS-PAGE gel and analyzed by Western blot using the indicated antibodies. *: non-specific bands

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