Journal: Cells

Article Title: Myeloid TM6SF2 Deficiency Inhibits Atherosclerosis

doi: 10.3390/cells11182877

Figure Lengend Snippet: TM6SF2 Contributes to Inflammatory Responses in Macrophages. ( A ) THP-1-derived macrophages were transfected with TM6SF2 siRNA or NT siRNA (20 nM) for 48 h and treated with oxLDL (100 µg/mL) for 4 h. ( B ) THP-1-derived macrophages were transfected with AdLacZ or AdTM6SF2 (100 MOI) for 48 h and treated with oxLDL (100 µg/mL) for 4 h. ( C ) BMDMs were isolated from control and Tm6sf2 mKO mice and treated with oxLDL (100 µg/mL) for 4 h. ( D ) BMDMs were isolated from wild type C57BL/6J mice and were transfected with AdLacZ or AdTM6SF2 (100 MOI) for 48 h and treated with oxLDL (100 µg/mL) for 4 h. mRNA expression of TNF-α , IL1β , and CCL2 were measured by qRT-PCR. n = 3 for each group. All data are presented as mean ± SEM, statistics by 2-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001 as indicated.

Article Snippet: After differentiation, macrophages were treated with 100 μg/mL oxLDL (360-31, by Lowry protein assay, Lee Biosolutions, Inc., Maryland Heights, MO, USA) for 48 h. Each sample was collected from an individual animal ( n = 4 for each group).

Techniques: Derivative Assay, Transfection, Isolation, Control, Expressing, Quantitative RT-PCR

Journal: Cells

Article Title: Myeloid TM6SF2 Deficiency Inhibits Atherosclerosis

doi: 10.3390/cells11182877

Figure Lengend Snippet: TM6SF2 contributes to foam cell formation. Macrophages were loaded with 5 μg/mL Dil-oxLDL at 37 °C for 40 min and collected for flow cytometry to detect the cholesterol uptake. Macrophages were loaded with 100 μg/mL oxLDL overnight at 37 °C and foam cell formation was determined by Oil Red O staining and the cellular cholesterol content (normalized to total protein). ( A , B ) THP-1-derived macrophages were transfected with NT siRNA or TM6SF2 siRNA (20 nM) for 48 h and treated with oxLDL (100 µg/mL) overnight. ( C , D ) THP-1-derived macrophages were transfected with AdLacZ or AdTM6SF2 (100 MOI) for 48 h and treated with oxLDL (100 µg/mL) overnight. ( E , F ) BMDMs were isolated from control and mKO mice and treated with oxLDL (100 µg/mL) overnight. ( G , H ) BMDMs were isolated from wild-type C57BL/6J mice and were transfected with AdLacZ or AdTM6SF2 (100 MOI) for 48 h and treated with oxLDL (100 µg/mL) overnight. n = 3 for flow cytometry, n = 5 for cholesterol contents detection. All data are presented as mean ± SEM, statistics by ordinary one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001 as indicated.

Article Snippet: After differentiation, macrophages were treated with 100 μg/mL oxLDL (360-31, by Lowry protein assay, Lee Biosolutions, Inc., Maryland Heights, MO, USA) for 48 h. Each sample was collected from an individual animal ( n = 4 for each group).

Techniques: Flow Cytometry, Staining, Derivative Assay, Transfection, Isolation, Control

Journal: Cells

Article Title: Myeloid TM6SF2 Deficiency Inhibits Atherosclerosis

doi: 10.3390/cells11182877

Figure Lengend Snippet: TM6SF2 Regulates ER stress-related Gene Expression in Macrophages. Macrophages were loaded with 100 µg/mL oxLDL overnight at 37 °C and mRNA expression of BIP , IRE1α , JNK , ASK1 and XBP1 were measured by qRT-PCR. ( A ) THP-1-derived macrophages were transfected with TM6SF2 siRNA or NT siRNA (20 nM) for 48 h and ( B ) THP-1-derived macrophages were infected with AdTM6SF2 or AdLacZ (100 MOI) for 48 h. ( C ) BMDMs were isolated from control and mKO mice and treated with oxLDL (100 µg/mL) overnight. ( D ) BMDMs were isolated from wild type C57BL/6J mice and were transfected with AdLacZ or AdTM6SF2 (100 MOI) for 48 h and treated with oxLDL (100 µg/mL) overnight. n = 3 for each group. All data are presented as mean ± SEM, statistics by 2way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001 as indicated.

Article Snippet: After differentiation, macrophages were treated with 100 μg/mL oxLDL (360-31, by Lowry protein assay, Lee Biosolutions, Inc., Maryland Heights, MO, USA) for 48 h. Each sample was collected from an individual animal ( n = 4 for each group).

Techniques: Gene Expression, Expressing, Quantitative RT-PCR, Derivative Assay, Transfection, Infection, Isolation, Control

Journal: Cells

Article Title: Myeloid TM6SF2 Deficiency Inhibits Atherosclerosis

doi: 10.3390/cells11182877

Figure Lengend Snippet: TM6SF2 increases p65 phosphorylation in THP-1-derived macrophages. ( A ) THP-1-derived macrophages were transfected with NT siRNA or TM6SF2 siRNA (20 nM) for 48 h and treated with oxLDL (100 µg/mL) for 1 h. ( B ) THP-1-derived macrophages were transfected with AdLacZ or AdTM6SF2 (100 MOI) for 48 h and treated with oxLDL (100 µg/mL) for 1 h. phospho-p65 and total p65 levels were determined by Western Blotting. Representative results and quantifications were shown. n = 3 for each group. All data are presented as mean ± SEM, statistics by Ordinary one-way ANOVA. * p < 0.05, ** p < 0.01 as indicated.

Article Snippet: After differentiation, macrophages were treated with 100 μg/mL oxLDL (360-31, by Lowry protein assay, Lee Biosolutions, Inc., Maryland Heights, MO, USA) for 48 h. Each sample was collected from an individual animal ( n = 4 for each group).

Techniques: Phospho-proteomics, Derivative Assay, Transfection, Western Blot

Journal: PLoS ONE

Article Title: 11β-Hydroxysteroid Dehydrogenase Type 1 Gene Knockout Attenuates Atherosclerosis and In Vivo Foam Cell Formation in Hyperlipidemic apoE −/− Mice

doi: 10.1371/journal.pone.0053192

Figure Lengend Snippet: Cytokine protein levels detected in cell culture media by protein multiplex analysis following overnight exposure of peritoneal foam cells to ∼30 µg/ml of oxidized LDL (Ox-LDL) vs. untreated controls. Peritoneal foam cells were harvested from Western diet-fed 11βHSD1 +/+ /apoE −/− (+/+) and 11βHSD1 −/− /apoE −/− (−/−) mice. G-CSF, KC, MCP1 and TNF-α levels were normalized to total cellular protein content. Cell culture media contained 200 nM 11-dehydrocorticosterone throughout the course of the experiment. Significance vs. control: *p≤0.05.

Article Snippet: Macrophages were incubated overnight in the presence or absence of ∼30 µg/ml copper-oxidized LDL (#BT-910, Biomedical Technologies, Stoughton, MA).

Techniques: Cell Culture, Multiplex Assay, Western Blot