Description of activities and resource inputs required for each activity.

<p>Description of activities and resource inputs required for each activity.</p

Program activities in each demand creation strategy.

<p>Program activities in each demand creation strategy.</p

Costs per intervention activity.

<p>Costs per intervention activity.</p

Costs per economic costing ingredient.

<p>Costs per economic costing ingredient.</p

Costs by thematic intervention activity.

<p>Costs by thematic intervention activity.</p

Additional file 1: of The Stepping Stones and Creating Futures intervention to prevent intimate partner violence and HIV-risk behaviours in Durban, South Africa: study protocol for a cluster randomized control trial, and baseline characteristics

Completed SPIRIT Guidelines for the Stepping Stones and Creating Futures Trial. (DOC 125 kb

Immunofluorescence cell imaging of PV1-infected HeLa cells in the presence and absence of BSO.

<p>(<b>A</b>) HeLa cells were infected with PV1-wt or PV1-BSOr (VP3 Q178L) at a moi of 10 at 37°C. After 4 hours incubation, the infected cells were probed for mature virus with A12 primary antibody (anti-PV human serum), which binds specifically to mature virus and provirions (Nihal Altan Bonnet, unpublished results), and Alexfluor 488-conjugated secondary antibody (green color). The localization of 3AB, a member of the replication complex, was determined in the same cell using 3AB mouse monoclonal antibody and Alexfluor 555-conjugated secondary antibody (red color). The scale is 5 µm. (<b>B</b>) Infected cells were probed for capsid precursors using VP3 polyclonal antibody and Alexfluor 488-conjugated secondary antibody (green color). The localization of 2C^ATPase, another nonstructural protein on the replication complex, was determined in the same cell using 2C mouse monoclonal antibody and Alexfluor 555-conjugated secondary antibody (red color).</p

Stepwise processing of the P1 capsid protein and maturation of the PV particle.

<p>Stepwise processing of the P1 capsid protein and maturation of the PV particle.</p

GSH protects PV1-wt and PV1-BSOr from heat inactivation.

<p>(<b>A</b>) Protection from heat inactivation of PV1-wt or PV1-BSOr by GSH. Purified PV1-wt or PV1-BSOr (VP3, Q178L) 3×10⁹ PFU was incubated <i>in vitro</i> in PBS either in the absence or presence of various amounts of GSH for 25 min at 48°C. The amount of virus remaining after the incubation was determined by plaque assays. The virus titers obtained after heating without GSH for both viruses were taken as “1”. (<b>B</b>) Protection from heat inactivation of PV1-wt or PV1-BSOr by reducing agents or GSSG. PV1-wt or PV1-BSOr 3×10⁹ PFU were incubated in PBS either in the absence or presence of various reducing agents (5 mM) or of GSSG (5 mM) (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004052#s4" target="_blank">Materials and Methods</a>). The virus titer obtained after heating without GSH for both viruses were taken as “1”.</p

Model for the role of GSH in the morphogenesis of C-cluster enteroviruses.

<p>Capsid precursor P1, in a complex with Hsp90, is processed by 3CD^pro yielding mature viral proteins VP0, VP1 and VP3, which spontaneously form 5S protomer. Under normal growth conditions, with ample supply of GSH, 5 protomers assemble into a pentamer (14S). The pentamers then either form an empty capsid or condense around the progeny RNA and form a provirion. The mature virus is derived from the provirion after the cleavage of VP0 to VP2 and VP4. When GSH levels are low, due to the presence of BSO, the pentamer is unstable. However, these unstable 14S pentamers can associate with RNA and form provirion-like particles, which can't mature into an infectious virus. The capsid of the BSOr mutants is more stable than that of the wt virus and requires much lower levels of GSH for the assembly of the precursors and the mature virus.</p