scd1 inhibitor a939572  (TargetMol)

Journal: Journal of Biochemistry

Article Title: Characterization of UGT8 as a monogalactosyl diacylglycerol synthase in mammals

doi: 10.1093/jb/mvae084

Figure Lengend Snippet: Membrane lipid saturation-induced UPR was suppressed in UGT8 KO HeLa cells. (A) Immunofluorescence of HeLa cells transiently expressing C-terminal FLAG-tagged mUgt8. Cells were immunostained with indicated antibodies. Scale bar, 10 μm. (B, C) The mRNA level of CHOP in WT or UGT8 KO (#1) HeLa cells treated with SCD1 inhibitor (100 nM) for 24 h (B) or tunicamycin (2.5 μg/ml) for 6 h (C) was quantified using qRT-PCR ( n = 3). (D–G) PERK phosphorylation of WT or UGT8 KO (#1) HeLa cells treated with SCD1 inhibitor (100 nM) for 24 h (D, E) or tunicamycin (2.5 μg/ml) for 6 h (F, G) were evaluated using immunoblotting (representative data of n = 2 out of n = 3 independent experiments). The ratio of phosphorylated PERK (pPERK) to total PERK was quantified (E, G) ( n = 3). (H–J) WT, UGT8 KO (#1) and UGT8 KO (#1) stably expressing mUgt8(WT)-FLAG or mUgt8(H358A)-FLAG HeLa cells were treated with SCD1 inhibitor (100 nM) for 24 h. PERK phosphorylation was evaluated using immunoblotting (F) (representative data of n = 2 out of n = 3 independent experiments). The ratio of phosphorylated PERK (pPERK) to total PERK was quantified (I) ( n = 3). The mRNA level of CHOP was quantified using qRT-PCR (J) ( n = 3). Mean ± SD. * P < 0.05, *** P < 0.001, **** P < 0.0001, ns; not significant, two-tailed unpaired t -test (B, C, E, G), one-way ANOVA with Dunnett’s multiple comparisons test (I, J).

Article Snippet: The UGT8 inhibitor (compound 19) and SCD1 inhibitor (A939572) were purchased from Cayman Chemicals.

Techniques: Membrane, Immunofluorescence, Expressing, Quantitative RT-PCR, Phospho-proteomics, Western Blot, Stable Transfection, Two Tailed Test

Journal: Regenerative Biomaterials

Article Title: Matrix stiffness boosts PDAC chemoresistance via SCD1-dependent lipid metabolic reprogramming

doi: 10.1093/rb/rbaf056

Figure Lengend Snippet: Matrix stiffness modulates SCD1 expression in Mia-PaCa2 cells. ( A ) Expression of SCD1 was markedly increased in tumor tissues ( n = 179) compared with those in normal tissues ( n = 171). ( B ) Representative immunohistochemistry staining images of SCD1 in tumor tissues and normal tissues. Scale bar: 20 μm. ( C ) The mRNA expression of SCD1. β-Actin as internal reference. ( D ) Representative SCD1 immunofluorescent images (red: SCD1; blue: DAPI). Scale bar: 20 μm for high magnifications and 50 μm for low magnifications. Zoomed-in areas were shown in white squares. ( E ) The protein expression and statistical analysis of SCD1. GAPDH as reference protein. ( F ) Representative SCD1 immunohistochemical staining images of Mia-PaCa2 cells encapsulated in the soft or stiff hydrogel after subcutaneous transplantation in vivo . Scale bar: 20 μm. Data were shown as the mean ± SD ( *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: For the stiff hydrogel group, PDAC cells were treated with 5 μM C75 (HY-12364; MCE) to inhibit the level of FA synthesis, handled with CAY10566 (SCD1 inhibitor, HY-15823; MCE) to inhibit the expression of SCD1, and 10 μM OA (HY-N1446; MCE) to increase FAs.

Techniques: Expressing, Immunohistochemistry, Staining, Immunohistochemical staining, Transplantation Assay, In Vivo

Journal: Regenerative Biomaterials

Article Title: Matrix stiffness boosts PDAC chemoresistance via SCD1-dependent lipid metabolic reprogramming

doi: 10.1093/rb/rbaf056

Figure Lengend Snippet: Matrix stiffness-induced fatty acid synthesis was suppressed in Mia-PaCa2 cells with pharmacological disruption of SCD1 activity. Cells were treated with DMSO or SCD1 inhibitor CAY10566 in the stiff group. ( A ) The mRNA expression of FASN, ACLY, ACACA and ACACB. β-Actin as internal reference. ( B ) Representative TEM images, red arrows indicate LDs. Scale bar: 2 μm for high magnifications and 5 μm for low magnifications. Zoomed-in areas were shown in white squares. ( C ) Cellular neutral lipids were determined by Nile Red and DAPI staining (red: LDs, blue: DAPI). ( D–F ) Representative immunofluorescent images of ACLY, FASN and ACC (purple: ACLY, FASN, ACC; blue: DAPI). Scale bar: 20 μm for high magnifications and 50 μm for low magnifications. Zoomed-in areas were shown in white squares. ( G ) Fluorescence quantitative analysis of Nile Red, ACLY, FASN and ACC. ( H ) The protein expressions and statistical analysis of ACLY, ACC and FASN. GAPDH as a reference protein. ( I ) Cellular TG content. Data were shown as the mean ± SD ( *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: For the stiff hydrogel group, PDAC cells were treated with 5 μM C75 (HY-12364; MCE) to inhibit the level of FA synthesis, handled with CAY10566 (SCD1 inhibitor, HY-15823; MCE) to inhibit the expression of SCD1, and 10 μM OA (HY-N1446; MCE) to increase FAs.

Techniques: Disruption, Activity Assay, Expressing, Staining, Fluorescence

Journal: Regenerative Biomaterials

Article Title: Matrix stiffness boosts PDAC chemoresistance via SCD1-dependent lipid metabolic reprogramming

doi: 10.1093/rb/rbaf056

Figure Lengend Snippet: The addition of OA restored the chemoresistance of SCD1 pharmacological disruption in Mia-PaCa2 cells. Cells were treated with DMSO, SCD1 inhibitor CAY10566 only or CAY10566 and OA in the stiff group. ( A ) The mRNA expression of ABCC1, ABCC3 and ABCC10. β-Actin as internal reference. ( B ) Representative immunofluorescent images of ABCC1 (top), and ABCC3 (bottom) (green: ABCC1, ABCC3; blue: DAPI). Scale bar: 20 μm for high magnifications and 50 μm for low magnifications. Zoomed-in areas were shown in white squares. ( C ) CCK-8 assay of gemcitabine sensitivity with increasing concentrations (0.01, 0.1, 1, 10, 100, 1000 μM, 5 replicates each) for 48 h (left). The statistical analysis of the IC 50 value (right). Data were shown as the mean ± SD ( *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: For the stiff hydrogel group, PDAC cells were treated with 5 μM C75 (HY-12364; MCE) to inhibit the level of FA synthesis, handled with CAY10566 (SCD1 inhibitor, HY-15823; MCE) to inhibit the expression of SCD1, and 10 μM OA (HY-N1446; MCE) to increase FAs.

Techniques: Disruption, Expressing, CCK-8 Assay

Journal: Regenerative Biomaterials

Article Title: Matrix stiffness boosts PDAC chemoresistance via SCD1-dependent lipid metabolic reprogramming

doi: 10.1093/rb/rbaf056

Figure Lengend Snippet: Matrix stiffness regulates SCD1 activation through Piezo1 in Mia-PaCa2 cells. Cells were treated with DMSO and Piezo1 agonist Yoda1 in the soft group or treated with DMSO and Piezo1 inhibitor GsMTx4 in the stiff group. ( A, B ) Representative immunofluorescent images of Piezo1 and SCD1 (red: Piezo1; purple: SCD1; blue: DAPI). Scale bar: 20 μm for high magnifications and 50 μm for low magnifications. Zoomed-in areas were shown in white squares. ( C, D ) Fluorescence quantitative analysis of Piezo1 and SCD1. ( E, F ) The mRNA expression of Piezo1 and SCD1. β-Actin as internal reference. ( G, H ) The protein expressions and statistical analysis of Piezo1 and SCD1. GAPDH as a reference protein. Data were shown as the mean ± SD ( *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: For the stiff hydrogel group, PDAC cells were treated with 5 μM C75 (HY-12364; MCE) to inhibit the level of FA synthesis, handled with CAY10566 (SCD1 inhibitor, HY-15823; MCE) to inhibit the expression of SCD1, and 10 μM OA (HY-N1446; MCE) to increase FAs.

Techniques: Activation Assay, Fluorescence, Expressing

Journal: Regenerative Biomaterials

Article Title: Matrix stiffness boosts PDAC chemoresistance via SCD1-dependent lipid metabolic reprogramming

doi: 10.1093/rb/rbaf056

Figure Lengend Snippet: Schematic overview showing that 3D matrix stiffness elicited lipid metabolism reprogramming to regulate PDAC chemoresistance via the mechanosensitive ion channel Piezo1-mediated SCD1, and the PI3K-Akt pathway participated in Piezo1-mediated SCD1 expression.

Article Snippet: For the stiff hydrogel group, PDAC cells were treated with 5 μM C75 (HY-12364; MCE) to inhibit the level of FA synthesis, handled with CAY10566 (SCD1 inhibitor, HY-15823; MCE) to inhibit the expression of SCD1, and 10 μM OA (HY-N1446; MCE) to increase FAs.

Techniques: Expressing