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. 2016 Jul 7;99(1):236–245. doi: 10.1016/j.ajhg.2016.05.026

Figure 3.

Figure 3

Subcellular Localization of the KLHL7 Mutants Found in CS/CISS1 Phenotype and Their Co-localization with CUL3

(A–X) HeLa cells cultured on coverslips were transfected with plasmids encoding either KLHL7:WT (A–F), KLHL7:p.Cys421Ser (G–L), KLHL7:p.Arg420Cys (M–R), KLHL7:p.Arg372Gln (S–X), and HA-CUL3 (kindly donated by Prof. Chiba). The cells were then fixed and stained with either anti-KLHL7 (Abnova Corp., cat# H00055975-B01P, RRID: AB_1576675; dilution 1:200) or anti-Myc (Abcam cat# ab18185, RRID: AB_444307; dilution 1:1,000) and anti-CUL3 antibodies (Bethyl cat# A301-109A, RRID: AB_873023; dilution 1:250). The nucleus was stained with DAPI. Confocal microscopy analysis shows KLHL7:WT and mutants in red and CUL3 in green. Scale bars represent 10 μm.

(Y) HeLa cells were transfected with plasmids encoding either KLHL7:WT, KLHL7:p.Cys421Ser, KLHL7:p.Leu341Trpfs∗9, KLHL7:p.Arg420Cys, KLHL7:p.Arg372Gln, and HA-CUL3, and the cell lysates were immunoblotted with anti-Myc, anti-HA, and anti-GAPDH. NT = not transfected.