Skip to main content
. 2020 Sep 9;107(4):753–762. doi: 10.1016/j.ajhg.2020.08.015

Figure 5.

Figure 5

Increased Partitioning of p.Lys33Glu and p.Arg42Trp Lamin B1 into the Cytosol

(A) Immunoblot analysis of lamin B1 and lamin A/C in nuclear extracts from LMNB1−/− HeLa cells transfected with WT and variant-bearing LMNB1 cDNAs (n = 4). Ponceau S staining is shown as a loading control.

(B) Immunoblot analysis of lamin B1 in whole-cell lysates of LMNB1−/− HeLa cells transfected with WT and variant-bearing LMNB1 cDNAs. The same transfected cells were subjected to subcellular fractionation to isolate nuclei and cytosol. These fractions were resolved by SDS-PAGE and analyzed by immunoblot. Representative images from four independent experiments are shown.

(C) Immunoblot analysis of LMNB1−/− HeLa whole-cell lysates and nuclear and cytosolic fractions from these cells following transfection with WT-, p.Arg29Trp-, and p.Arg42Trp-encoding cDNAs. GAPDH and lamin A/C blots are shown below (n = 3).

(D) Collective quantification by densitometry of the ratio of lamin B1 to lamin A/C in nuclear extracts shown in (A)–(C). Values were normalized to the ratio observed with WT LMNB1 cDNA (ratio was set at 1.0 for each condition). Error bars denote standard deviation of the mean; p values were determined via Student’s t test (∗∗p < 0.01; ∗∗∗p < 0.001).