Journal: International Journal of Biological Sciences

Article Title: The Interaction of Calcium-Sensing Receptor with KIF11 Enhances Cisplatin Resistance in Lung Adenocarcinoma via BRCA1/cyclin B1 pathway

doi: 10.7150/ijbs.92046

Figure Lengend Snippet: KIF11 interacted with CaSR and was upregulated in DDP-resistant LUAD cells. Mass spectrometry (MS) was used to identify proteins (A-C) , CaSR-interacting proteins (A) , KIF11-interacting proteins (B) , and proteins interacting with CaSR or KIF11 are selected based on two screening criteria: The protein was not pulled down by IgG or the protein had less than 2 folds increase in detection in IgG compared to CaSR or KIF11 (C) . Protein-protein interaction maps between CaSR and KIF11 (D) , CaSR is depicted in dark blue, and KIF11 is depicted in cyan. The corresponding binding points are indicated as clubbed structures of the corresponding colors. And western blotting results showed clear detection of KIF11 in the CaSR-interacting proteins (E) and CaSR in the KIF11-interacting proteins (F) . KIF11 was highly expressed in LUAD (G) . Kaplan-Meier plots showed the correlation of KIF11 with overall survival in LUAD (H) , including the patient underwent chemotherapy (I) .

Article Snippet: A kinesin family member 11 (KIF11) inhibitor (EG5 Inhibitor V, trans-24; T11155) was purchased from TargetMol (Boston, MA, USA).

Techniques: Mass Spectrometry, Binding Assay, Western Blot

Journal: International Journal of Biological Sciences

Article Title: The Interaction of Calcium-Sensing Receptor with KIF11 Enhances Cisplatin Resistance in Lung Adenocarcinoma via BRCA1/cyclin B1 pathway

doi: 10.7150/ijbs.92046

Figure Lengend Snippet: Downregulation of KIF11 inhibited the growth of DDP-resistant LUAD cells. A549-DDP and H1299-DDP cells were transfected with siRNA targeting KIF11 (siKIF11-1, siKIF11-2) or non-targeting siRNA (si-NC). After KIF11 knockdown, the clonogenic proliferation of cisplatin-resistant cells was disrupted (A) . The proliferation ability of A549-DDP and H1299-DDP were inhibited and the panels show the relative cell viability at the 48 h (B) . Western blotting results showed the changes on protein levels of KIF11, CaSR, cyclin B1, CDK1 and BRCA1 (C) . Data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 and ns, not significant.

Article Snippet: A kinesin family member 11 (KIF11) inhibitor (EG5 Inhibitor V, trans-24; T11155) was purchased from TargetMol (Boston, MA, USA).

Techniques: Transfection, Knockdown, Western Blot

Journal: International Journal of Biological Sciences

Article Title: The Interaction of Calcium-Sensing Receptor with KIF11 Enhances Cisplatin Resistance in Lung Adenocarcinoma via BRCA1/cyclin B1 pathway

doi: 10.7150/ijbs.92046

Figure Lengend Snippet: Inhibition of KIF11 reduced cisplatin resistance in DDP-resistant LUAD cells. The clonogenic proliferation of DDP-resistant cells was disrupted by the KIF11 inhibitor (A) . A549-DDP cells and H1299-DDP cells were treated with 10 µM cisplatin and 1µM KIF11 inhibitor, respectively, or in combination. Colony formation assay was performed to analyze the cells, and the right panel quantifies the relative number of colonies formed after 14 days. The sensitivity of A549-DDP (B and C) and H1299-DDP (D and E) cells to cisplatin was affected by the KIF11 inhibitor. The protein levels of KIF11, CaSR, cyclin B1, CDK1 and BRCA1 were detected by western blotting (F) . The schematics illustrate the proposed mechanism of CaSR activates the cisplatin resistance in LUAD (G) . Data are showed as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 and ns, not significant.

Article Snippet: A kinesin family member 11 (KIF11) inhibitor (EG5 Inhibitor V, trans-24; T11155) was purchased from TargetMol (Boston, MA, USA).

Techniques: Inhibition, Colony Assay, Western Blot

Journal: The Journal of Physiological Sciences : JPS

Article Title: Involvement of receptor-interacting protein 140 in estrogen-mediated osteoclasts differentiation, apoptosis, and bone resorption

doi: 10.1007/s12576-016-0447-2

Figure Lengend Snippet: Involvement of RIP140 in estrogen-inhibited osteoclast differentiation and bone resorption. a Verified RIP140 knockdown effect by Western blot in lentivirus-mediated transduction of primary culture osteoclasts precursors. b Verified RIP140 knockdown effect by qRT-PCR in lentivirus-mediated transduction of primary culture osteoclasts precursors. n = 3, **p < 0.01. c TRAP staining of osteoclasts precursors with or without 10−8 M estrogen and transfected with lentivirus-shRNA-targeting RIP140. d Summarized data showed that the depletion of RIP140 significantly attenuated the inhibitor effect of estrogen on osteoclastogenesis. n = 3, *p < 0.05. **p < 0.01. e Osteoclast activity was measured by release of europium-labeled collagen measured by florescence. n = 3, *p < 0.05. **p < 0.01. f Pit formation assay of osteoclasts precursors with or without 10−8 M estrogen and transfection with Lentivirus-shRNA-targeting RIP140. g Summarized data showed that the depletion of RIP140 attenuated the inhibitor effect of estrogen on osteoclasts bone resorption activity. n = 3, *p < 0.05. **p < 0.01. h–j qRT-PCR analysis of osteoclasts marker gene in osteoclasts precursors with or without estrogen and transfection with Lentivirus-shRNA-targeting RIP140. Con control, NC negative control, E2 17β-estradiol

Article Snippet: Mature osteoclasts were infected with the FasL shRNA lentiviral particles (Santa Cruz, CA, USA) for 24 h. Furthermore, NC cells were transduced with control shRNA lentiviral particles (Santa Cruz, CA, USA).

Techniques: Knockdown, Western Blot, Transduction, Quantitative RT-PCR, Staining, Transfection, shRNA, Activity Assay, Labeling, Tube Formation Assay, Marker, Control, Negative Control

Journal: The Journal of Physiological Sciences : JPS

Article Title: Involvement of receptor-interacting protein 140 in estrogen-mediated osteoclasts differentiation, apoptosis, and bone resorption

doi: 10.1007/s12576-016-0447-2

Figure Lengend Snippet: Estrogen induced osteoclast apoptosis by increasing RIP140 expression through the Fas/FasL pathway. a Regulation of caspase-3 activity by estrogen. Osteoclasts were treated for 12 h with 10−8 M estrogen. n = 3, **p < 0.01. b The levels of Bcl-2 and Bax protein from osteoclasts treated with estrogen were analyzed by Western blot. c Western-blot analysis of RIP140 protein expression in osteoclasts treated with 10−8 M estrogen for 12 h. d qRT-PCR analysis of RIP140 mRNA expression in osteoclasts treated with 10−8 M estrogen for 12 h. n = 3, **p < 0.01, *p < 0.05. e The effects of estrogen and Lentivirus-shRNA-targeting RIP140 on caspase-3 activity in osteoclasts. n = 3, **p < 0.01, *p < 0.05. f The levels of Bcl-2 and Bax protein from osteoclasts treated with estrogen and Lentivirus-shRNA-targeting RIP140 were analyzed by Western blot. g qRT-PCR analysis of FasL mRNA expression in osteoclasts treated with 10−8 M estrogen for 12 h. n = 3, **p < 0.01. h qRT-PCR analysis of Fas mRNA expression in osteoclasts treated with 10−8 M estrogen for 12 h. n = 3. i Western-blot analysis of RIP140 protein expression in osteoclasts treated with estrogen and Lentivirus-shRNA-targeting FasL. j qRT-PCR analysis of RIP140 mRNA expression in osteoclasts treated with estrogen and Lentivirus-shRNA-targeting FasL. n = 3, **p < 0.01, *p < 0.05. Con control, NC negative control, E2 17β-estradiol. k A schematic diagram of RIP140-involved pathways on estrogen-regulated osteoclastogenesis, osteoclast apoptosis, and bone resorption

Article Snippet: Mature osteoclasts were infected with the FasL shRNA lentiviral particles (Santa Cruz, CA, USA) for 24 h. Furthermore, NC cells were transduced with control shRNA lentiviral particles (Santa Cruz, CA, USA).

Techniques: Expressing, Activity Assay, Western Blot, Quantitative RT-PCR, shRNA, Control, Negative Control

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Clinico-histopathological characteristics of the 136 cutaneous melanomas according to CD33, VISTA and PD-1 expression.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Correlations among CD33, VISTA, and PD-1 expression.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Expression of CD33 and VISTA in cutaneous melanoma. ( A ) High intratumoral expression (immunohistochemical score of 3, red color, cytoplasmic) of CD33 in acral melanoma (× 100). ( B ) High intratumoral expression (score 3, red color, cytoplasmic) of VISTA in nodular melanoma (× 100). ( C ) High peritumoral expression (score 3, red color, cytoplasmic) of CD33 in acral melanoma (× 100). ( D ) High peritumoral expression (score 3, red color, cytoplasmic) of VISTA in nodular melanoma (× 200). ( E ) Dual immunofluorescence staining of ( A ) CD33 and VISTA (CD33, red, cytoplasmic; VISTA, green, cytoplasmic) and ( F ) PD-1 and VISTA (PD-1, red, cytoplasmic; VISTA, green, cytoplasmic). Double-positive cells are shown in yellow (white arrow). VISTA, V-domain Ig suppressor of T-cell activation; PD-1, programmed cell death protein-1.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing, Immunohistochemical staining, Immunofluorescence, Staining, Activation Assay

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Patient survival according to CD33 and VISTA expression. ( A ) OS according to CD33 expression. ( B ) OS according to VISTA expression. ( C ) Worse OS in patients with high expression of both CD33 and VISTA. ( D ) OS according to the degree of CD33 and VISTA expression ( E ) Worse OS in cases with dual expression of VISTA and PD-1 among patients with high expression of PD-1. VISTA, V-domain Ig suppressor of T-cell activation; PD-1, programmed cell death protein-1.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing, Activation Assay

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Univariate and multivariate analyses of OS and PFS.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing