Journal: Scientific Reports

Article Title: Epigallocatechin-3-gallate inhibits H 2 O 2 -induced apoptosis in Mouse Vascular Smooth Muscle Cells via 67kD Laminin Receptor

doi: 10.1038/s41598-017-08301-6

Figure Lengend Snippet: Effect of EGCG on H 2 O 2 -induced changes in 67LR. ( A ) Immunocytochemistry analysis was performed using anti-67LR antibody and Alexa Flouor-488 conjugated secondary antibody(green). The nuclei were stained with DAPI(blue). VSMCs were pretreated with 10, 50, 100, or 150 μM EGCG for 2 h, followed by treatment with 200 μM H 2 O 2 for 30 min. ( B ) Western blot showing the expression of 67LR after treatment with EGCG and H 2 O 2 . Results were expressed as Mean ± S.D. from 5 independent experiments. *P < 0.05 vs. H 2 O 2 treatment group, # p < 0.05 vs. control group.

Article Snippet: Primary antibodies against caspase-8 (#13423-1-AP), caspase-9 (#10380-1-AP), Bcl-2 (#12789-1-AP), Bax (#50599-2-Ig), CathepsinD (#21327-1-AP), 67LR (#14533-1-AP), GAPDH (#10494-1-AP) and β-actin (#20536-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: Immunocytochemistry, Staining, Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: Epigallocatechin-3-gallate inhibits H 2 O 2 -induced apoptosis in Mouse Vascular Smooth Muscle Cells via 67kD Laminin Receptor

doi: 10.1038/s41598-017-08301-6

Figure Lengend Snippet: The efficiency of shRNA silencing of 67LR. ( A ) Transfection efficiency in 293 T cells. ( B ) Western blot analysis showing the expression of 67LR after silencing by shRNA plasmid. *p < 0.05 vs. Scramble group.

Article Snippet: Primary antibodies against caspase-8 (#13423-1-AP), caspase-9 (#10380-1-AP), Bcl-2 (#12789-1-AP), Bax (#50599-2-Ig), CathepsinD (#21327-1-AP), 67LR (#14533-1-AP), GAPDH (#10494-1-AP) and β-actin (#20536-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: shRNA, Transfection, Western Blot, Expressing, Plasmid Preparation

Journal: Scientific Reports

Article Title: Epigallocatechin-3-gallate inhibits H 2 O 2 -induced apoptosis in Mouse Vascular Smooth Muscle Cells via 67kD Laminin Receptor

doi: 10.1038/s41598-017-08301-6

Figure Lengend Snippet: The protective effects of EGCG on H 2 O 2 -induced apoptosis following shRNA silencing of 67LR. ( A ) Western blot analysis showing the expression levels of caspase-3, caspase-8 and caspase-9 following shRNA silencing of 67LR, and subsequent pretreatment with 10, 50, 100, or 150 μM EGCG for 2 h, followed by treatment with 200 μM H 2 O 2 for 30 min. ( B ) Western blot analysis showing the expression levels of Bcl-2, Bax and Cathepsin D following shRNA silencing of 67LR, and subsequent pretreatment with 10, 50, 100, or 150 μM EGCG for 2 h, followed by treatment with 200 μM H 2 O 2 for 30 min. β-actin and GAPDH were used as internal controls. Results were expressed as Mean ± S.D. from 5 independent experiments. *P < 0.05 vs. H 2 O 2 treatment group, # p < 0.05 vs. control group.

Article Snippet: Primary antibodies against caspase-8 (#13423-1-AP), caspase-9 (#10380-1-AP), Bcl-2 (#12789-1-AP), Bax (#50599-2-Ig), CathepsinD (#21327-1-AP), 67LR (#14533-1-AP), GAPDH (#10494-1-AP) and β-actin (#20536-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: shRNA, Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: Epigallocatechin-3-gallate inhibits H 2 O 2 -induced apoptosis in Mouse Vascular Smooth Muscle Cells via 67kD Laminin Receptor

doi: 10.1038/s41598-017-08301-6

Figure Lengend Snippet: The 67LR shRNA knockdown sequences.

Article Snippet: Primary antibodies against caspase-8 (#13423-1-AP), caspase-9 (#10380-1-AP), Bcl-2 (#12789-1-AP), Bax (#50599-2-Ig), CathepsinD (#21327-1-AP), 67LR (#14533-1-AP), GAPDH (#10494-1-AP) and β-actin (#20536-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: shRNA, Knockdown

Journal: Cell Death & Disease

Article Title: A small molecule binding HMGB1 inhibits caspase-11-mediated lethality in sepsis

doi: 10.1038/s41419-021-03652-5

Figure Lengend Snippet: A Confocal microscopy of mouse peritoneal macrophages incubated for 4 h with DQ ovalbumin (10 μg/mL; red) alone or together with LPS-binding protein (10 μg/mL), or together with HMGB1(10 μg/mL) in the absence or presence of FeTPPS (1 μM) or PIX (1 μM) for 4 h, then stained with DAPI (blue). Scale bar: 10 μm. B Western blots for cathepsin D, Na + -K + -ATPase, Lamp1, Rab7 in the cytosolic fraction from vehicle-treated or LBP (5 μg/mL) or HMGB1 (5 μg/mL) with or without FeTPPS(1 μM) or PIX (1 μM)-treated mouse peritoneal macrophages. C Flow cytometry of mouse peritoneal macrophages stained with acridine orange and then treated for 4 h with LBP (10 μg/mL) or HMGB1 (10 μg/mL) with or without FeTPPS (1 μM) Numbers above bracketed lines indicate percent cells with loss of lysosomal staining with acridine orange (excitation, 488 nm; emission, 650–690 nm). D Whole-cell patch-clamp recording of HMGB1 in the absence or presence of FeTPPS (1 μM) induced inward current across the cytoplasmic membrane in proximity to the patch-clamp of HEK293 cells at acidic conditions (pH = 5.0). Graphs show the mean ± SD of technical replicates and are representative of at least three independent experiments.

Article Snippet: Recombinant mouse LPS-binding protein (LBP) (6635-LP-025/CF) was from R&D Systems Inc.

Techniques: Confocal Microscopy, Incubation, Binding Assay, Staining, Western Blot, Flow Cytometry, Patch Clamp, Membrane

Journal: Shock

Article Title: Lipopolysaccharide-Induced Cytokine Secretion from In Vitro Mouse Slow and Fast Limb Muscle

doi: 10.1097/shk.0000000000001891

Figure Lengend Snippet: FIG. 1. Representative cytokines secreted from isolated SOL in response to 1 mg/mL LPS dissolved in Krebs buffer (n ¼ 8 per group), Stats are Wilcoxon Signed ranks for matched pairs *P < 0.05, ** P < 0.01. T0 ¼ the beginning of LPS exposure, T1 ¼ 1 h after LPS exposure, T2 ¼ 2 h after LPS.

Article Snippet: Tests of SOL muscles were repeated in response to 1 mg/mL LPS in the presence of 1% C57BL/6 cytokine-free plasma (IGMS-C57-N, Innovative Research, Novi, Mich), 1 mg/mL LPS þ 1 mg/mL LPS-binding protein (LBP; 6635-LP/CF, R&D Systems, Minneapolis, Minn) or Krebs buffer alone.

Techniques: Isolation

Journal: Shock

Article Title: Lipopolysaccharide-Induced Cytokine Secretion from In Vitro Mouse Slow and Fast Limb Muscle

doi: 10.1097/shk.0000000000001891

Figure Lengend Snippet: FIG. 2. Representative cytokine secretory responses in isolated mouse SOL and mouse extensor digitorum longus (EDL) in response to 1 mg/mL LPS in Krebs buffer þ 1% serum. Stats used Wilcoxon- Signed ranks for matched pairs from time 0 to T1, and T1 to T2. ** ¼ P < 0.01, *** P < 0.001, n ¼ 10 for SOL, n ¼ 17 for EDL. T0 ¼ the beginning of LPS exposure, T1 ¼ 1 h after LPS exposure, T2 ¼ 2 h after LPS.

Article Snippet: Tests of SOL muscles were repeated in response to 1 mg/mL LPS in the presence of 1% C57BL/6 cytokine-free plasma (IGMS-C57-N, Innovative Research, Novi, Mich), 1 mg/mL LPS þ 1 mg/mL LPS-binding protein (LBP; 6635-LP/CF, R&D Systems, Minneapolis, Minn) or Krebs buffer alone.

Techniques: Isolation

Journal: Shock

Article Title: Lipopolysaccharide-Induced Cytokine Secretion from In Vitro Mouse Slow and Fast Limb Muscle

doi: 10.1097/shk.0000000000001891

Figure Lengend Snippet: FIG. 4. Comparisons of total cytokine secretion (log scale) of SOL vs. EDL at T2. Muscles were incubated in Krebs þ 1% plasma. T2 ¼ accumulated cytokines at the end of the 2 h exposure to LPS. Means SD, n ¼ 11 for SOL, n ¼ 16 for EDL. Comparisons are two sample t for parametric and Mann–Whitney for nonparametric samples. * ¼ P < 0.05, ** P < 0.01

Article Snippet: Tests of SOL muscles were repeated in response to 1 mg/mL LPS in the presence of 1% C57BL/6 cytokine-free plasma (IGMS-C57-N, Innovative Research, Novi, Mich), 1 mg/mL LPS þ 1 mg/mL LPS-binding protein (LBP; 6635-LP/CF, R&D Systems, Minneapolis, Minn) or Krebs buffer alone.

Techniques: Muscles, Incubation, Clinical Proteomics, MANN-WHITNEY