ZFIN Image: Sofou et al., 2021, Figure EV3

Image

Figure Caption

Figure EV3 Analysis of HOPS/CORVET subunits in fibroblasts (related to Fig 3)

A, B

Effect of proteasome inhibition on VPS16 and VPS33A. (A) Immunoblots for VPS16 and VPS33A and quantifications of protein levels in lysates of fibroblasts treated with MG132 for 30 or 120 min, respectively. VPS16 band densities were quantified in an extended area (indicated by red boxes) to capture the presence of a possible truncated protein with a molecular weight of 87 kDa (wild‐type VPS16 is 95 kDa). Stars denote unspecific bands. (B) Quantifications of (A) normalized to control conditions and represented as mean ± SEM for VPS16 (n = 3 biological replicates) or mean for VPS33A (n = 2 biological replicates).

C–E

Rescue of cellular phenotype by VPS^N52K. (C) Representative immunoblots of fibroblast lysates using antibodies against VPS16, VPS33A, and actin. Stars denote unspecific bands. (D) Levels of VPS16 and VPS33A quantified in patient cells transduced with control or VPS16^N52K‐expressing lentiviruses and normalized to levels of actin (n = 3). Data represented as mean ± SEM; ***P < 0.001 by unpaired Student’s t‐tests. (E) Cross‐species sequence alignment of VPS16 residues surrounding the asparagine in position 52; (bottom row) asterisks, colons, and periods indicate residues that are fully, strongly, or weakly conserved, respectively.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ EMBO Mol. Med.