Journal: ACS Omega

Article Title: NUSAP1 Drives Endometrial Carcinoma via IP3R Phosphorylation, Calcium Dyshomeostasis, and ER-Stress-Mediated Apoptosis

doi: 10.1021/acsomega.5c05485

Figure Lengend Snippet: NUSAP1 mediated the phosphorylation of IP3R. (A–D) Expression levels of NUSAP1, p-IP3R, and IP3R in HEC-1-B and ISK cells were detected by Western blotting after the depletion or overexpression of NUSAP1. n = 3, ** P < 0.01, *** P < 0.001 vs NC-siRNA, ### P < 0.001 vs NC-OE. (D–G) Immunofluorescence of NUSAP1 (green), p-IP3R (green), IP3R (green), and DAPI (blue) of following knockdown or overexpression of NUSAP1. n = 3, *** P < 0.001 vs NC-siRNA, ### P < 0.001 vs NC-OE. siRNA, small-interfering RNA; OE, overexpression.

Article Snippet: The primary antibodies used were as follows: NUSAP1 (1:5000, 12024-1-AP, 750 μg/mL, Proteintech), IP3R (1:1000, DF3000, 1 mg/mL, Affinity), P-IP3R (1:1000, DF2999, Affinity), Bax (1:2000, 50599-2-IG, 800 μg/mL, 1 mg/mL, Proteintech), Bcl-2 (1:1000, 610538,BD), IRE1 (1:1000, 27528-1-AP, 500 μg/mL, Proteintech), PERK (1:1000, 20582-1-AP,800 μg/mL, Proteintech), PINK1 (1:1000, bsm-51265M, 1 mg/mL, Bioss),Parkin (1:1000, bs-23687R, 1 mg/mL, Bioss).

Techniques: Phospho-proteomics, Expressing, Western Blot, Over Expression, Immunofluorescence, Knockdown, Small Interfering RNA

Journal: ACS Omega

Article Title: NUSAP1 Drives Endometrial Carcinoma via IP3R Phosphorylation, Calcium Dyshomeostasis, and ER-Stress-Mediated Apoptosis

doi: 10.1021/acsomega.5c05485

Figure Lengend Snippet: NUSAP1-IP3R interaction regulated calcium dynamics and ER stress to modulate cell fate. (A, B) Co-IP assay verified the binding between NUSAP1 and IP3R in HEC-1-B and ISK cells. (C–F) Efficiency of IP3R knockdown was validated using Western blotting in HEC-1-B and ISK cells. n = 3, *** P < 0.001 vs NC-siRNA. (G–I) Fluo-4 AM green fluorescence staining was used to detect Ca 2+ levels of HEC-1-B and ISK cells following the transfection of IP3R-siRNA, NUSAP1-OE + IP3R-siRNA, and NUSAP1-siRNA + IP3R-siRNA (scale bar, 100 μm), n = 3, *** P < 0.001vs IP3R-siRNA, ## P < 0.01, ### P < 0.001 vs IP3R-siRNA, small-interfering RNA; OE, overexpression; Ca 2+ , calcium ion.

Article Snippet: The primary antibodies used were as follows: NUSAP1 (1:5000, 12024-1-AP, 750 μg/mL, Proteintech), IP3R (1:1000, DF3000, 1 mg/mL, Affinity), P-IP3R (1:1000, DF2999, Affinity), Bax (1:2000, 50599-2-IG, 800 μg/mL, 1 mg/mL, Proteintech), Bcl-2 (1:1000, 610538,BD), IRE1 (1:1000, 27528-1-AP, 500 μg/mL, Proteintech), PERK (1:1000, 20582-1-AP,800 μg/mL, Proteintech), PINK1 (1:1000, bsm-51265M, 1 mg/mL, Bioss),Parkin (1:1000, bs-23687R, 1 mg/mL, Bioss).

Techniques: Co-Immunoprecipitation Assay, Binding Assay, Knockdown, Western Blot, Fluorescence, Staining, Transfection, Small Interfering RNA, Over Expression