Figure S2.

arl13 cilia accumulate AMPK and PLC. (A and B) Bar graphs showing the relative levels of PLD (A) and FAP12 (B) in the strains indicated based on Western blot quantifications. The number of biological repeats, i.e., cilia preparations, is indicated; error bars show the standard deviation. (C) Western blot characterizing the anti-FAP12 antibody using isolated cilia of control (WT, g1) and a fap12 insertional mutant (CLiP strain LMJ.RY0402.206664), which carries an insertion in the fourth exon deleting four bases. Anti-IC2 and anti-IFT81 were used as loading controls. The fap12 mutant will be characterized elsewhere. (D) Schematic presentation of the ARL13-NG expression vector. (E) Western blot of isolated cilia from control (g1, WT) and arl13 strains probed with antibodies against FMG1, PKD2, CAH6, ARL13, and IC2, as a loading control. The analysis is based on several membranes with equal loading; the lanes stained with anti-ARL13 and anti-IC2 were also used for the Western blot shown in Fig. 2 A. (F) Western blot of isolated cilia from control (g1, WT) and arl13 strains probed with GT335 to detect polyglutamylated protein, anti-ARL13, anti-FAP12, anti-PLD and anti-IC2, as a loading control. The numbers indicate band intensities (WT = 1), the stars indicate that the values were adjusted for loading based on the IC2 signal. One of two biological replicates is shown. (G) Western blot of isolated cilia from control (g1, WT), arl13 and bbs4-1 strains probed with antibodies against PLC and IC2, as a loading control. Antibodies to PLC were characterized by Awasthi et al. (2012). (H) Western blot of isolated cilia from control (g1, WT) and arl13 probed with antibodies against AMPK, and IC2, as a loading control. (I) Western blot of isolated cilia from control (g1, WT), bbs4-1 arl13, bbs4-1 arl13 ARL13-NG, and arl13 ARL13-NG strains probed with antibodies against BBS4, ARL13, and IC2, as a loading control. Source data are available for this figure: SourceData FS2.