Journal: International Journal of Molecular Sciences

Article Title: Biosynthesized Calcium Peroxide Nanoparticles as a Multifunctional Platform for Liver Cancer Therapy

doi: 10.3390/ijms26104696

Figure Lengend Snippet: CaO 2 NPs as nanocarriers promote apoptosis in liver cancer cells. ( A ) The Cell Counting Kit (CCK-8) assay of MHCC 97-H cells. Group (1): the cells were treated with CaO 2 NPs (150 mg/L), DTX (37.5 mg/L), or DTX@CaO 2 NPs (containing 37.5 mg/L DTX) and tested after incubation for 48 h. Group (2): The cells were treated with CaO 2 NPs (300 mg/L), DTX (75 mg/L), or DTX@CaO 2 NPs (containing 75 mg/L DTX) and tested after incubation for 48 h. The bars represent the results from three independent experiments ( n = 3); *** p < 0.001. ( B ) The flow cytometry analysis using Annexin V/PI staining. MHCC 97-H cells were treated with 150 mg/L CaO 2 NPs, 37.5 mg/L DTX, or DTX@CaO 2 NPs (containing 37.5 mg/L DTX) for 24 h. ( C ) The quantitative analysis of the apoptotic cells in B ( n = 3); * p < 0.033, *** p < 0.001.

Article Snippet: The apoptosis assay kit was purchased from Multi Sciences (Hangzhou, China).

Techniques: Cell Counting, CCK-8 Assay, Incubation, Flow Cytometry, Staining

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 3. Effects of hypoxia on CXCL6 and CXCL2 gene expression in co-culture systems. (A) The difference of CXCL6 gene expression between censored (n ¼ 95) and relapse/refractory (n ¼ 247) patients in TARGET database. (B to E) The gene expression of CXCL6 and CXCL2 of MSCs and HUAECs in the co-culture systems were measured by the co-culture/mono culture ratio. (B and D) MSC þ THP-1/MSC; (C and E) HUAEC þ THP-1/HUAEC. *P < 0.05, **P < 0.01, ***P < 0.001, and “ns” for no statistical significance. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Gene Expression, Co-Culture Assay

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 4. CXCL6 and CXCL2 content in the supernatant of hypoxia and THP-1 co-culture systems. (A and B) CXCL6 content in supernatant. (C and D) CXCL2 content in supernatant. *P < 0.05, **P < 0.01, and ***P < 0.001. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Co-Culture Assay

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 5. The effects of CXCL6 and CXCL2 on the proliferation and migration of MSCs and HUAECs. (A and B) The effects of CXCL6 (A) and CXCL2 (B). Among them, (a and b) described the effects of cytokines on MSCs and (c and d) described the effects of cytokines on HUAECs. (a and c) showed the effects of different concentrations of cytokines on target cells. (b and d) showed the time-dependent effect of cytokines on target cells. (C) The chemotaxis of CXCL6 and CXCL2 to MSCs and HUAECs respectively. The concentration of CXCL6 was 50 ng/mL and of CXCL2 was 100 ng/mL. Scale ¼ 100 mm. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Migration, Chemotaxis Assay, Concentration Assay

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 6. Expression of cleaved-caspase3 in MSCs and HUAECs. (A) Expression of cleaved-caspase-3 (green fluorescence) measured by immunofluorescence method increased significantly in hypoxia (1% O2) for 48 h, but decreased after CXCL6 was supplied. The reversion depended on the concentration of CXCL6. MSCs were pictured at 20 water objective and HUAECs were pictured at 20 air objective. The heatmap represented the change of average fluorescence intensity in each group. (B) Western blot method verified the above results. The histograms (MSCs on the left and HUAECs on the right) represented the relative gray value of each group. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Expressing, Fluorescence, Immunofluorescence, Concentration Assay, Western Blot

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 7. Angiogenesis ability of vascular endothelial cells authenticated in vitro. (A) HUAECs in standard culture conditions, CoCl2 simulate hypoxia and hypoxia plus CXCL6 or CM conditions in vitro were used to simulated angiogenesis. HUVECs tubule forming experiment was taken as positive control. (B to E) Quantitative comparison of the number of master junction and meshes as well as total meshes area and total segments length. Scale ¼ 200 mm. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: In Vitro, Positive Control, Comparison

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 8. Correlation between CXCL6 gene expression and HIF-1a gene expression in AML patients. The original data were downloaded from Cbioporta (https:// www.cbioportal.org/datasets), n ¼ 323. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Gene Expression

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 9. Regulation of CXCL6 expression by HIF-1a. (A) MSCs or HUAECs were treated with CoCl2 (200 mmol/L), with or without BAY87-2243 (1 mmol/L), and the protein expression of HIF-1a was detected by Western blot method after 24 or 48 h. (B) The relative expression of HIF-1a in three independent experiments was statistically analyzed. The x-axis coordinates represented CoCl2()/BAY(), CoCl2(þ)/BAY(), CoCl2(þ)/BAY(þ), respectively. (a) MSC–24 h; (b) MSC–48 h; (c) HUAEC–24 h; (d) HUAEC–48 h. (C) MSCs and HUAECs were cultured for 24 or 48 h under the condition of 1% O2, with or without BAY87-2243 (1 mmol/L). The relative expression of CXCL6 mRNA was detected by real-time PCR. (a) MSC–24 h; (b) MSC–48 h; (c) HUAEC–24 h; (d) HUAEC–48 h. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Expressing, Western Blot, Cell Culture, Real-time Polymerase Chain Reaction

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 11. CXCL6 activate mTOR in MSCs and HUAECs. (A) CoCl2 (200 mmol/L) was used to treat MSCs or HUAECs for 24 h with or without CXCL6 of 50 ng/Ml. (B) The statistical results of three groups of independent experiments, **P < 0.01. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques:

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Hypoxia-CXCL6 axis affects arteriolar niche remodeling in acute myeloid leukemia.

doi: 10.1177/1535370220960675

Figure Lengend Snippet: Figure 10. Negative feedback regulation of CXCL6 on HIF-1a. (A) MSCs and HUAECs were cultured for 24 or 48 h under 1% O2 condition with or without CXCL6 (50 ng/mL). The relative mRNA expression of HIF-1a was detected by real-time PCR. (a) MSC–24 h; (b) MSC–48 h; (c) HUAEC–24 h; (d) HUAEC–48 h. (B) MSCs or HUAECs were treated with CoCl2, with or without CXCL6 (50 ng/mL). After 24 or 48 h, the expression of HIF-1a protein was detected by Western blot method. (C) The relative expression of HIF-1a protein in three independent experiments was statistically analyzed, and the x-axis coordinates were respectively representative CoCl2()/CXCL6(), CoCl2(þ)/CXCL6(), CoCl2(þ)/CXCL6(þ), (a) MSC–24 h; (b) MSC–48 h; (c) HUAEC–24 h; (d) HUAEC–48 h. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and “ns” for no statistical significance. (A color version of this figure is available in the online journal.)

Article Snippet: The cytokine levels in the supernatant of each group were respectively detected by the human CXCL6 ELISA kit (BOSTER, Wuhan, China) and human CXCL2 ELISA Kit (MULTI SCIENCES, Hangzhou, China) according to the manufacturer’s protocol.

Techniques: Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Western Blot