Journal: Cell Death and Differentiation

Article Title: Suppressor of hepatocellular carcinoma RASSF1A activates autophagy initiation and maturation

doi: 10.1038/s41418-018-0211-7

Figure Lengend Snippet: RASSF1A interacts with HDAC6 to enhance acetylation of α-Tubulin and co-localizes with acetylated microtubules. a , b Representative immunoblot ( a ) and quantification ( b ) showing levels of acetylated α-tubulin (Ac-α-Tub) in liver tissues from 4-month-old wild-type and RASSF1A −/− mice. c , d Representative immunoblot ( c ) and quantification ( d ) showing levels of acetylated α-tubulin in hepatocytes isolated from wild-type and RASSF1A −/− mice. e , f Representative immunoblot ( e ) and quantification ( f ) showing levels of acetylated α-tubulin in HeLa cells transiently transfected with control plasmid, plasmid expressing HA-RASSF1A (HA-RA), and plasmid expressing HA-RASSF1A∆ (HA-RA∆). g , h Representative images ( g ) and quantification ( h ) showing the immunostaining intensities of acetylated α-tubulin in cells similar to those in e . Bar = 20 µm. Red, HA-RA or HA-RA∆; green, acetylated a-tubulin; and yellow, colocalization. i , j Representative immunoblot ( i ) and quantification ( j ) showing the impact of RASSF1A on levels of HDAC6 in cells similar to those in e . k , l Representative immunoblots showing levels of Flag-HDAC6 coimmunoprecipitated with endogenous RASSF1A from HeLa cells ( k ) or exogenous RASSF1A in 293T cells ( l ) transiently overexpressing Flag-HDAC6. m Representative immunoblots showing levels of RA-HA or RA-RA∆ coimmunoprecipitated with Flag-HDAC6 in 293T cells. n Representative immunoblot showing levels of GFP fused fragments of RASSF1A coimmunoprecipitated with HDAC6 from lysates of HeLa cells transiently expressing GFP fused RASSF1A constructs

Article Snippet: Plasmids encoding Myc-LC3 (#24919), HDAC6 (#30482) and P27 (#14049) were purchased from Addgene.

Techniques: Western Blot, Isolation, Transfection, Control, Plasmid Preparation, Expressing, Immunostaining, Construct