polyclonal rabbit anti scd1  (Cell Signaling Technology Inc)

Journal: Journal of Ovarian Research

Article Title: Stearoyl-CoA desaturase 1 inhibition induces ER stress-mediated apoptosis in ovarian cancer cells

doi: 10.1186/s13048-024-01389-1

Figure Lengend Snippet: SCD1 is highly expressed in ovarian cancer tissues compared to normal tissues. ( A, B ) The microarray data for SCD1 mRNA expression in normal ovarian surface epithelial (NOSE) and epithelial ovarian cancer (EOC) tissue samples was obtained from Gene Expression Omnibus (GEO) database, including ( A ) GSE14407 and ( B ) GSE26712. ( C ) Representative IHC images showing SCD1 expression pattern in adjacent normal tissues and ovarian cancer tissues in TMA sections. SCD1 expression was scored as 0, 1+, 2+, or 3 + based on staining intensity. ( D ) Different IHC score of SCD1 protein expression between adjacent normal and ovarian cancer tissues. Values are presented as means ± SEM (** p < 0.01; *** p < 0.001)

Article Snippet: IHC staining was conducted using a rabbit polyclonal antibody against SCD1 (1:200, bs-3787R, Bioss, Woburn, MA).

Techniques: Microarray, Expressing, Staining

Journal: Journal of Ovarian Research

Article Title: Stearoyl-CoA desaturase 1 inhibition induces ER stress-mediated apoptosis in ovarian cancer cells

doi: 10.1186/s13048-024-01389-1

Figure Lengend Snippet: SCD1 expression is significantly elevated in EOC cell lines compared to NOSE cell lines. ( A ) The mRNA expression levels of SCD1 in EOC cell lines and NOSE cell lines. SCD1 mRNA levels were analyzed by qRT-PCR analysis and normalized to GAPDH mRNA levels. ( B ) The protein expression levels of SCD1 in EOC cell lines and NOSE cell lines were detected by western blot analysis. ( C ) SCD1 protein levels were quantified by densitometry using ImageJ software followed by normalization to GAPDH protein levels. Data are presented as the mean ± SEM of three independent experiments

Article Snippet: IHC staining was conducted using a rabbit polyclonal antibody against SCD1 (1:200, bs-3787R, Bioss, Woburn, MA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Software

Journal: Journal of Ovarian Research

Article Title: Stearoyl-CoA desaturase 1 inhibition induces ER stress-mediated apoptosis in ovarian cancer cells

doi: 10.1186/s13048-024-01389-1

Figure Lengend Snippet: SCD1 inhibition reduces cancer cell proliferation without cytotoxic effect on normal cells. ( A ) PA-1 and SKOV-3 cells were transfected with SCD1 siRNA (100 nM) or scrambled siRNA (100 nM) as a negative control. After 72 h, the ablation of SCD1 protein was determined by western blot analysis. GAPDH was used as a loading control. ( B ) PA-1 and SKOV-3 cells were transfected with SCD1 siRNA (100 nM). At 72 h post-transfection, cell viability was analyzed using MTT assay. ( C ) PA-1 and SKOV-3 cells were treated with various concentrations (0, 5, 10, 20, 50, and 100 nM) of CAY10566 or DMSO (solvent control) for 24–48 h. Cell viability was measured by MTT assay. ( D ) Bright field microscopy images of patient-derived ovarian cancer organoids A209 (top) and A220 (bottom) after 28 days of culture. Scale bar = 500 μm. ( E ) Ovarian cancer organoids A209 and A220 were treated with varying concentrations (0, 10, 20, 50, and 100 nM) of CAY10566 or DMSO (solvent control) for 24–48 h. Cell viability was assessed using the organoid MTT assay. ( F ) CAY10566 were treated in NOSE cell lines (IOSE385 and SNU3236) and peripheral blood mononuclear cells (PBMCs) with different concentrations of CAY10566. After 24–48 h, cell viability was examined by MTT assay. All data were described as mean ± SEM of three independent experiments (** p < 0.01; *** p < 0.001; **** p < 0.0001)

Article Snippet: IHC staining was conducted using a rabbit polyclonal antibody against SCD1 (1:200, bs-3787R, Bioss, Woburn, MA).

Techniques: Inhibition, Transfection, Negative Control, Western Blot, Control, MTT Assay, Solvent, Microscopy, Derivative Assay

Journal: Journal of Ovarian Research

Article Title: Stearoyl-CoA desaturase 1 inhibition induces ER stress-mediated apoptosis in ovarian cancer cells

doi: 10.1186/s13048-024-01389-1

Figure Lengend Snippet: MUFA/SFA ratio alteration induced by SCD1 inhibition leads to ER stress-mediated apoptosis. ( A ) PA-1 and SKOV-3 cells were treated with the IC50 value of CAY10566 for PA-1 cells (20 nM) for 48 h. Desaturase activity of SCD1 was estimated as the ratios of palmitoleic acid (C16:1n7) to palmitic acid (C16:0) and oleic acid (C18:1n9c) to stearic acid (C18:0) using gas chromatography. The statistical analysis was conducted using a two-way ANOVA followed by Šídák’s multiple comparisons test. ( B ) PA-1 cells were transfected with SCD1 siRNA (100 nM) or treated with CAY10566 (20 nM) for 48 h. The percentage of apoptotic cells was determined by flow cytometry analysis using Annexin V-FITC and PI staining. ( C ) PA-1 cells were transfected with SCD1 siRNA (100 nM) or treated with CAY10566 (20 nM) for 48 h. The expression levels of apoptosis marker proteins, PARP and cleaved caspase-3, were detected by western blot analysis. GAPDH was used as a loading control. ( D ) PA-1 cells were transfected with SCD1 siRNA (100 nM) or treated with CAY10566 (20 nM) for 48 h. The expression levels of ER stress marker proteins, p-PERK, IRE1α, ATF4, and CHOP, were examined by western blot analysis. GAPDH was used as a loading control. All data were described as mean ± SEM of three independent experiments (** p < 0.01; *** p < 0.001; **** p < 0.0001)

Article Snippet: IHC staining was conducted using a rabbit polyclonal antibody against SCD1 (1:200, bs-3787R, Bioss, Woburn, MA).

Techniques: Inhibition, Activity Assay, Gas Chromatography, Transfection, Flow Cytometry, Staining, Expressing, Marker, Western Blot, Control

Journal: Journal of Ovarian Research

Article Title: Stearoyl-CoA desaturase 1 inhibition induces ER stress-mediated apoptosis in ovarian cancer cells

doi: 10.1186/s13048-024-01389-1

Figure Lengend Snippet: Addition of exogenous oleic acid rescued ER stress-mediated apoptosis triggered by SCD1 inhibition. ( A ) PA-1 cells were treated with CAY10566 (20 nM) with oleic acid-BSA (10 mM) or fatty acid-free BSA (10 µM) as vehicle control for 48 h. Cell viability was analyzed by MTT assay. Results were presented as the percentage of total cell number compared to DMSO-treated control. ( B ) PA-1 cells were treated with CAY10566 (20 nM) with or without oleic acid-BSA (10 µM) for 48 h. The percentage of apoptotic cells was calculated by flow cytometry analysis using Annexin V-FITC and PI staining. ( C ) Inhibition of CAY10566-mediated PARP and caspase-3 cleavage by supplementation with oleic acid. PA-1 cells were treated with CAY10566 (20 nM) with or without oleic acid-BSA (10 mM) for 48 h. The expression levels of cleaved PARP and caspase-3 were assessed by western blot analysis. GAPDH was used as a loading control. ( D ) PA-1 cells were treated with CAY10566 (20 nM) with or without oleic acid-BSA (10 µM) for 48 h. The expression levels of P-PERK, IRE1α, ATF4, and CHOP were determined by western blot analysis. GAPDH was used as a loading control. All data were described as mean ± SEM of three independent experiments (* p < 0.05; ** p < 0.01; *** p < 0.001)

Article Snippet: IHC staining was conducted using a rabbit polyclonal antibody against SCD1 (1:200, bs-3787R, Bioss, Woburn, MA).

Techniques: Inhibition, Control, MTT Assay, Flow Cytometry, Staining, Expressing, Western Blot