Journal: BMC Infectious Diseases

Article Title: Comparison of apoptosis in human primary pulmonary endothelial cells and a brain microvascular endothelial cell line co-cultured with Plasmodium falciparum field isolates

doi: 10.1186/s12879-017-2552-0

Figure Lengend Snippet: Cytoadherence and apoptosis of HLEC and hCMEC/D3 mediated by P. falciparum infected red blood cell

Article Snippet: A suspension of 300 μl of mature pRBC at 6% parasitaemia and 2% haematocrit (approximately 4 × 10 6 schizonts for 63 × 10 6 uninfected RBC) in complete RPMI culture medium without bicarbonate was added to each well containing confluent HLEC and hCMEC/D3 monolayers from Labtek in duplicate, and incubated for 1 h at 37 °C.

Techniques: Infection

Journal: BMC Infectious Diseases

Article Title: Comparison of apoptosis in human primary pulmonary endothelial cells and a brain microvascular endothelial cell line co-cultured with Plasmodium falciparum field isolates

doi: 10.1186/s12879-017-2552-0

Figure Lengend Snippet: PRBC-mediated EC apoptosis. a ) PRBC-mediated EC apoptosis by direct contact with and without cytoadherence The ability of field isolates to induce EC apoptosis was the same for hCMEC/D3 (9 apoptogenic PRBCs) and HLEC (4 apoptogenic PRBCs) ( p > 0.05) and the mean optical density was also similar ( p > 0.05); b ) PRBC-mediated EC apoptosis by release of soluble factors. hCMEC/D3 were more susceptible to apoptosis mediated by soluble factors (11 apoptogenic PRBCs) than were HLEC (4 apoptogenic PRBCs) ( p < 0.05) but intensity (mean OD ratio) of this apoptosis was higher with hCMEC/D3 ( p < 0.01). P. falciparum field isolates trigger cerebral cells apoptosis than lung cells ( p < 0.01)

Article Snippet: A suspension of 300 μl of mature pRBC at 6% parasitaemia and 2% haematocrit (approximately 4 × 10 6 schizonts for 63 × 10 6 uninfected RBC) in complete RPMI culture medium without bicarbonate was added to each well containing confluent HLEC and hCMEC/D3 monolayers from Labtek in duplicate, and incubated for 1 h at 37 °C.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: The YAP–TEAD4 complex promotes tumor lymphangiogenesis by transcriptionally upregulating CCBE1 in colorectal cancer

doi: 10.1016/j.jbc.2023.103012

Figure Lengend Snippet: YAP/TAZ and TEAD4 promote tumor lymphangiogenesis in CRC in vitro.A, in CRC, high nuclear TEAD4 expression is correlated with an increased lymph node metastasis rate compared with that of CRC tumors with low TEAD4 expression. The original immunoreactive score and clinical data were obtained from our previous study (20). ∗∗∗p < 0.001 by the chi-squared test. B, HLEC tube formation assay with conditioned medium from TEAD4 knockdown HCT116 cells. HLECs were cultured with conditioned medium from the indicated HCT116 cells. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by Student’s t test. C and D, tube formation and wound healing assays of HLECs cultured with conditioned medium from the indicated SW837 cells (C) and CAFs (D). The scale bars for the tube formation assay represent 100 μm. The scale bars for the wound healing assay represent 200 μm. ∗p < 0.05, ∗∗p < 0.01 by Student’s t test. CAF, cancer-associated fibroblast; CRC, colorectal cancer; HLEC, human lymphatic endothelial cell.

Article Snippet: HLEC (#2500, lot no.19415) were purchased from ScienCell and cultured in Endothelial Cell Medium (#1001, ScienCell).

Techniques: In Vitro, Expressing, Tube Formation Assay, Cell Culture, Wound Healing Assay

Journal: The Journal of Biological Chemistry

Article Title: The YAP–TEAD4 complex promotes tumor lymphangiogenesis by transcriptionally upregulating CCBE1 in colorectal cancer

doi: 10.1016/j.jbc.2023.103012

Figure Lengend Snippet: JQ1 inhibits lymphangiogenesis by suppressing the expression of CCBE1 in CRC.A, JQ1 decreased the mRNA level of CCBE1 in HCT116 and SW837 cells and primary cancer-associated fibroblasts derived from two different CRC tissues. Cells were treated with JQ1 (1 μM) or DMSO for 24 h, and the mRNA level of CCBE1 was then determined by quantitative. ∗∗p < 0.01, ∗∗∗∗p < 0.0001 by Student’s t test. B, knockdown of BRD2/3/4 by siRNAs decreased the mRNA level of CCBE1 in HCT116 and SW837 cells. Cells were transfected with the indicated siRNA for 72 h, and the mRNA level of CCBE1 was then determined by quantitative PCR. ∗∗∗∗p < 0.0001 by Student’s t test. C, Western blot analysis of CCBE1 protein levels in supernatants from the indicated SW837 cells treated with JQ1 (1 μM) or DMSO for 24 h. D, Western blot analysis of pro-VEGFC and mature VEGFC protein levels in the indicated mixed conditioned medium. Conditioned medium from the indicated treated SW837 cells was mixed with conditioned medium from full-length VEGFC-expressing 293T cells (1:1), incubated overnight and analyzed by Western blotting. E, Western blot analysis of CCBE1 protein levels in supernatants from the indicated treated SW837 cells overexpressing CCBE1 and treated with JQ1 (1 μM) or DMSO for 24 h. F, Western blot analysis of pro-VEGFC and mature VEGFC protein levels in the indicated mixed conditioned medium. Conditioned medium from the indicated treated stable SW837 cells was mixed with conditioned medium from full-length VEGFC-expressing 293T cells (1:1), incubated overnight, and analyzed by Western blotting. Ponceau S staining was used to control for equal loading. G, tube formation and wound healing assays of HLECs cultured with conditioned medium from the indicated SW837 cells. To exclude the effect of JQ1 on HLECs, SW837 cells were treated with JQ1 (1 μM) or DMSO for 24 h and were then washed and cultured with fresh culture medium without JQ1 for another 24 h. This conditioned medium without JQ1 was collected for the assay with HLECs. The scale bars for the tube formation assay represent 100 μm. The scale bars for the wound healing assay represent 200 μm. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by Student’s t test. H, immunohistochemical analysis of CCBE1 and mLyve-1 (lymphatic vessel number) in the indicated treated stable HCT116 cell line–derived xenografts in mouse footpads. Representative images of mLyve-1(+) lymphatic vessels and CCBE1 protein expression in the same field are shown. Black arrows: mLyve-1(+) lymphatic vessels. Stars: CRC cells. The scale bars represent 20 μm. ∗p < 0.05, ∗∗p < 0.01, by Student’s t test. I, immunohistochemical analysis of CCBE1 and mLyve-1 in seven CRC patient-derived xenografts treated with JQ1 (50 mg/kg). Representative images are shown. The scale bars represents 20 μm. Black arrows: mLyve-1(+) lymphatic vessels. PDX samples treated with JQ1 had lower CCBE1 scores in tumor cells/stroma and lower lymphatic vessel densities than those treated with DMSO. ∗p < 0.05 by the Mann–Whitney U test for CCBE1 scores; ∗∗p < 0.01 by paired Student’s t test for mLyve-1(+) lymphatic vessel counts. CRC, colorectal cancer; DMSO, dimethyl sulfoxide; HLEC, human lymphatic endothelial cell. PDX, patient-derived xenograft.

Article Snippet: HLEC (#2500, lot no.19415) were purchased from ScienCell and cultured in Endothelial Cell Medium (#1001, ScienCell).

Techniques: Expressing, Derivative Assay, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Incubation, Staining, Cell Culture, Tube Formation Assay, Wound Healing Assay, Immunohistochemical staining, MANN-WHITNEY