Lovastatin treatment of DENV-infected cells does not inhibit ER rearrangement.

<p>Mock-infected 2fTGH cells were treated with 10 µM lovastatin for 12 h and were stained intracellularly for (A) SREBP-2 (green) and HMGCR (red). (B) Cells were also stained for DENV proteins E (green) and NS3 (red). 2fTGH cells were infected with DENV-2 and mock-treated for 12 h. Cells were fixed and stained intracellularly for (C) SREBP-2 (magenta) and (D) HMGCR (magenta), followed by DENV E (green) and NS3 (red) staining. Nuclear staining (blue) was performed using DAPI. DENV-infected 2fTGH cells were treated with 10 µM lovastatin at the time of infection and incubated for 12 h. Cells were fixed and stained as described above for (E) SREBP2 (magenta) and (F) HMGCR (magenta) and DENV E (green) and NS3 (red). Nuclear staining (blue) was performed using DAPI, and images were acquired as previously described. Total magnification 630×.</p

V_Ms3_Dermis mono Tx steadystate left

File type: The repository includes FSC 3.0 files that were exported from Diva software (BD Biosciences) and can be opened with FlowJo software (TreeStar). In some cases, an updated compensation matrix is included for a specific experiment that should be applied to the FCS files within the FlowJo software. Fluorescent channels also indicate the specifically stained markers

II_Epidermis 24h primary DENV Ifnar

File type: The repository includes FSC 3.0 files that were exported from Diva software (BD Biosciences) and can be opened with FlowJo software (TreeStar). In some cases, an updated compensation matrix is included for a specific experiment that should be applied to the FCS files within the FlowJo software. Fluorescent channels also indicate the specifically stained markers

DENV induces rearrangement of the ER early after infection.

<p>2fTGH cells were (A) mock-infected or (B–E) infected with DENV-2 for 12 h and stained intracellularly for UPR markers and viral proteins E and NS3. (A) Mock-infected 2fTGH cells were stained with mouse MAbs against DENV E (4G2; green) and NS3 (E1D8; red) directly conjugated to Alexa Fluor® 488 and Alexa Fluor® 594, respectively. Nuclear staining (blue) was performed using DAPI stain. Intracellular staining of DENV-infected cells was carried out as in (A); in addition primary antibodies specific for cellular proteins (B) GRP78, (C) GRP94, (D) XBP1 and (E) ATF6 (magenta) were used, followed by secondary antibodies conjugated to Alexa Fluor® 647. The multiple-exposure images were captured using 63X/1.25 Plan-Neofluar DIC oil objective on Zeiss 510 LSM using bandpass filter sets appropriate for DAPI, Alexa Fluor® 488, Alexa Fluor® 594 and Alexa Fluor® 647. For individual images, the additional channels were turned OFF post-acquisition using the Zeiss operating software. Co-localization is observed in the “Merged” image as a yellow hue. Merged and DIC images represent a single image with all detector channels ON. Total magnification 630×. (F) Western blot analysis of GRP78 over a 12-h course of DENV infection. Actin was used as a loading control, and expression of DENV NS1 was used to confirm viral infection.</p

III_Dermis 24h ADE DENV Ifnar

File type: The repository includes FSC 3.0 files that were exported from Diva software (BD Biosciences) and can be opened with FlowJo software (TreeStar). In some cases, an updated compensation matrix is included for a specific experiment that should be applied to the FCS files within the FlowJo software. Fluorescent channels also indicate the specifically stained markers

IV_Exp 48 & 72h Compensation Matrix

File type: The repository includes FSC 3.0 files that were exported from Diva software (BD Biosciences) and can be opened with FlowJo software (TreeStar). In some cases, an updated compensation matrix is included for a specific experiment that should be applied to the FCS files within the FlowJo software. Fluorescent channels also indicate the specifically stained markers

cho-bies

chovy nsticks of wood feathered out with a knife for kindling.YesUsed I and SupNot usedNot usedchoby, chopy, chuffy, shovy, MOP, MOP-HEAD, SHAVINGSChecked by Cathy Wiseman on Fri 26 Jun 201

Annual Collection of Serological Samples, Managua, Nicaragua, 2005

<div><p>The collection of samples, monitored here by Eva Harris (right), is part of the UC Berkeley/SSI pediatric dengue cohort study in Managua, Nicaragua. Objectives of this study include investigating the natural history and risk factors for dengue virus transmission and disease, obtaining biological samples in parallel with clinical and epidemiological information, and establishing a site for a future phase III trial of a safe tetravalent dengue vaccine.</p><p>(Photo: Alejandro Belli)</p></div

DENV-induced ER rearrangement and expansion is independent of the XBP-1 pathway.

<p>XBP1^+/+ (A–D) and XBP1^−/− (E–H) MEF cells were infected with DENV-2 for 12 h and stained intracellularly for cellular proteins using antibodies against (A and E) GRP78, (B and F) GRP94, (C) XBP1, and (G) ATF6 (magenta), followed by secondary antibodies conjugated to Alexa Fluor® 647. DENV proteins were detected with mouse MAbs against DENV E (green) and NS3 (red) directly conjugated to Alexa Fluor® 488 and Alexa Fluor® 594, respectively. Nuclear staining (blue) was performed using DAPI stain. Images were acquired and processed as described in Fig. 1. Total magnification 630×.</p

I_Dermis steadystate Mono stain WT

File type: The repository includes FSC 3.0 files that were exported from Diva software (BD Biosciences) and can be opened with FlowJo software (TreeStar). In some cases, an updated compensation matrix is included for a specific experiment that should be applied to the FCS files within the FlowJo software. Fluorescent channels also indicate the specifically stained markers