Fig. 3

a GST pull-down analyses of purified GST-NudCL2 with His-CP110. b GST pull-down analyses of purified GST-LC3 with His-NudCL2. c In vitro protein interaction of purified GST-LC3 with His-CP110 and His-NudCL2. d, e Wild-type (WT) and NudCL2 knockout (KO-1) MEF cells were applied for co-IP analysis with anti-CP110 or anti-LC3 antibodies, respectively. 3% of total input is shown. f–j WT and NudCL2 KO MEF cells were transfected with the indicated plasmids, cultured with or without serum for 24 h, and processed for the following analyses. Western blot analysis showed the expression of the indicated proteins (f, g). β-actin was used as a loading control. Immunofluorescence analyses were carried out by using anti-CEP164 and anti-CP110 antibodies (h). Scale bar, 2 µm. The percentage of cells with CP110 dots at mother centrioles was calculated (i, j). k Schematic diagram shows the possible LIR motifs (W/F/YxxL/I/V) of NudCL2. The point mutations (depicted in red) in four putative LIR motifs are defined as M1, M2, M3, and M4, respectively. l MEF cells transfected with the indicated plasmids were subjected to co-IP experiments. 3% of total input is shown. m GST pull-down analysis of purified GST-LC3 with His-NudCL2 or His-M2. n In vitro protein interaction of purified GST-LC3 with His-CP110 and His-NudCL2 or His-M2. Quantitative data are expressed as the means ± SD (at least three independent experiments). n, sample size. *P < 0.05 and **P < 0.01; Student’s t-test.