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Blood, Vol. 113, Issue 6, 1278-1286, February 5, 2009
Cell adhesion through  V-containing integrins is required for efficient HIV-1 infection in macrophages
Blood Ballana et al.
113: 1278
Supplemental materials for: Ballana et al
Files in this Data Supplement:
- Figure S1. siRNA mediated silencing of integrin alphaV did not change HIV-1 receptor/coreceptor expression (JPG, 88 KB)
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Flow cytometry histograms showing alphaV, CD4 and CXCR4 staining two days after siRNA transfection. Cells treated with siAlphaV#1 (red line), siAlphaV#2 (blue line) or siNon-targeting (brown line) are shown compared to mock-treated cells (solid black line) and unstained negative control (dotted black line). A representative experiment is shown.
- Figure S2. qRT-PCR confirmed specific inhibition of alphaV and beta5 mRNAs by their corresponding siRNAs (JPG, 28 KB)
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mRNA levels of alphaV (black bars) and beta5 integrin (white bars) were quantified by RT-PCR, relativized by ΔCt to GAPDH mRNA levels and normalized to the mock-treated control. Mean±SD of two independent experiments is shown.
- Figure S3. siRNA-treated HeLa cells partially recovered alphaV surface expression 5 days post-tranfection (JPG, 38.7 KB)
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Flow cytometry histograms showing alphaV staining two and five days after siRNA transfection. Cells treated with siAlphaV#1 (red line) or siAlphaV#2 (blue line) are shown compared to mock-treated cells (solid black line) and unstained negative control (dotted black line). A representative experiment is shown.
- Figure S4. siRNA or S36578 treatment did not change cell viability in HeLa cells or MDM (JPG, 34.7 KB)
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(A) HeLa cells transfected with siRNA were counted 5 days after transfection. Number of cells was relativized to the mock-treated cells and expressed as mean±SD of at least 3 independent experiments (B) Uninfected HeLa cells treated with S36578 and AZT were counted 5 days after seeding. Number of cells was relativized to the cells treated without drug (ND) and expressed as mean±SD of at least 3 independent experiments (C) Absolute absorbance (550/620) of MTT assay performed in uninfected MDM and treated for 7 days with indicated compounds. A representative experiment is shown.
- Figure S5. HIV-1 replication under adhesive and non-adhesive conditions in HeLa cells (JPG, 38.9 KB)
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(A) Absolute number of recovered cells 5 days after S36578 treatment of HeLa cells cultured under adhesive (black bars) or non-adhesive (white bars) conditions. A representative experiment is shown. (B) Absolute values of beta-galactosidase assay (HIV replication) in HeLa cells cultured under adhesive (upper panel) or non-adhesive (lower panel) conditions 5 days after HIV-1 infection. A representative experiment is shown.
- Figure S6. HIV-1 replication under adhesive and low-adhesive conditions in MDM (JPG, 91.9 KB)
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(A) Absolute values of recovered p24 (pg/ml) in cell supernatants of MDM from three different donors seven days post-infection with R5 strain HIV-1 BaL, cultured under adhesive (upper panel) or low-adhesive (lower panel) conditions. Low-adhesive cultures were infected with 2 fold virus compared to their corresponding adhesive MDM. Mean±SD of three replicates is shown. (B) Absolute absorbance (550/620) of MTT assay performed in uninfected MDM cultured in adhesive (black bars) or low-adhesive (white bars) conditions and treated for 7 days with indicated compounds. A representative experiment is shown.
- Figure S7. AlphaV staining of HeLa cells and MDM cultured under adhesive and non/low-adhesive conditions (JPG, 36.6 KB)
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Representative flow cytometry histograms showing alphaV staining (red line) compared with unstained samples (black dotted line) in HeLa and MDM cells cultured under non/low-adhesive conditions.
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