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Costume designer Ane Crabtree gives the rundown on dressing Margaret Atwood’s dystopia.
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Courtesy of Hulu.

In the Republic of Gilead—which once was Cambridge, Massachusetts, until the state was overthrown by a theocratic, totalitarian regime—women are defined solely by their assigned roles and the men who govern them. Wives, Daughters, Aunts, Marthas, Econowives, and Handmaids live to serve their male counterparts; their free will and reproductive rights have been stripped in an attempt to restore life and order to the mostly infertile republic.

Most post-1980s high school students—those whose libraries didn’t ban the book, anyway—will recognize Gilead from The Handmaid’s Tale , a 1985 work of speculative fiction by famed writer and environmental activist Margaret Atwood . If the dystopian tale was required reading then, its new small-screen adaptation—coming to Hulu April 26th—feels like required viewing now, and not just for teens. Costume designer Ane Crabtree , tasked with bringing Gilead’s dystopian dress to life, agrees, admitting that her creative process was particularly emotional and charged because the show’s filming schedule aligned with the results of the 2016 presidential election.

Margaret Atwood Ane Crabtree
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“By November, when the world changed in our own personal Gilead on this side of the pond, we were well into the story, and every day the script was mirroring what was happening in the States,” says Crabtree. “The world changed in a matter of days with the commander that we have, and it was a blessing and a curse to work on this show and creatively build for that. Because I think artistic torment is actually a blessing. It was painful some days, and beautiful on other days. That’s the only way to describe it.”

SRα and the 12-kDa subunit of the signal peptidase complex (SPC12), representing complexes involved in targeting of nascent chains to the ER and cleavage of signal peptides, respectively, were more uniformly distributed among the subfractions from both liver and adrenal (see Figs. 5 and 7 ). However, αTRAP, one of four subunits of the TRAP complex, which colocalizes with native ribosome-channel complexes, was localized primarily in the ribosome-bearing subfractions from both adrenal and liver (see Figs. 5 and 7 ). Little αTRAP was detected in the smooth microsomes obtained from either organ.

Sec62 and Sec63, mammalian homologs of proteins essential for posttranslational translocation in yeast, were present in both the rough and smooth microsomal subfractions from liver and adrenal (data not shown).

We examined several proteins involved in folding of newly synthesized polypeptides and ER quality control, generally considered to be distributed throughout the ER: two lumenal proteins, BiP and GRP94, and one membrane protein, calnexin (see Table 1 ). In liver microsomes, particularly those of the guinea pig, BiP and GRP94 were in higher concentration in the rough microsomes (see Fig. 7 ). In adrenal microsomal subfractions, their concentrations in smooth microsomes were equal to or slightly greater than in rough microsomes ( Fig. 5 ). Calnexin was present in all microsomal subfractions from liver but did not show a consistent differential pattern of distribution (see Fig. 7 ). In adrenal microsomes, the levels of calnexin were higher in the smooth microsomes ( Fig. 5 ).

Having found high levels in adrenal smooth microsomes of all translocon subunits and associated elements examined, except for ribosomal protein and αTRAP, it seemed important to confirm that preferential localization in smooth microsomes of enzymes involved in sterol and steroid synthesis was retained in these subcellular fractions. Therefore, we examined the levels of HMGR and two enzymes involved in steroid synthesis, 3βHSD and 17α-hydroxylase, a cytochrome P450 (CYP17). All of these enzymes were in highest concentration in the adrenal smooth microsomes ( Fig. 5 ). None were detectable in corun rough microsomes from liver or pancreas, although HMGR was present in liver smooth and intermediate microsomes (data not shown).

To determine whether the translocon components and steroidogenic enzymes have a similar distribution in situ , immunocytochemistry was performed on isolated adrenocortical cells. These cells retain their abundant SER in vitro , particularly in the presence of ACTH ( 74 ). Both CYP17 and Sec61were distributed fairly evenly throughout the tubular ER network of the isolated cells ( Fig. 6 , A and B, respectively). Corun controls showed minimal staining ( Fig. 6C ). Little change in the distribution of CYP17 and Sec61 was detected in ACTH-treated cells at this resolution (data not shown). However, in some cells, hotspots of intense labeling occurred for both proteins, which seemed to be more abundant in ACTH-treated cells (data not shown). More detailed analysis will be required to see whether these correspond to the organized arrays of SER observed in these cells with the electron microscope ( 4 6 ).

Fox Theatre
Fox Theatre
660 Peachtree Street NE / Phone: (404) 881-2100 Box Office: (855) 285-8499
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