Journal: Molecular Medicine

Article Title: The therapeutic potential of Apigenin in amyotrophic lateral sclerosis through ALDH1A2/Nrf2/ARE signaling

doi: 10.1186/s10020-024-00977-7

Figure Lengend Snippet: Effect of APG on viability and apoptosis in NSC34 neuronal cells. A CCK8 assay to evaluate the effects of different doses of APG on the viability of NSC34 cells in vitro. B Flow cytometry analysis of apoptosis in NSC34 cells harboring the SOD1*G93A mutation treated with APG. C Western blot analysis of apoptosis-related proteins (Bcl-2, caspase-3, Bax) in SOD1*G93A mutant NSC34 cells treated with APG. D Flow cytometry assessment of ROS levels in SOD1*G93A mutant NSC34 cells under APG treatment. E - H ELISA for quantification of OS markers (MDA, 4-HNE, SOD, ALDH) in SOD1*G93A mutant NSC34 cells treated with APG. I qRT-PCR analysis of ALDH1A2 mRNA levels, and (J) Western blot analysis of ALDH1A2 protein levels in APG-treated SOD1*G93A mutant NSC34 cells. * P < 0.05, ** P < 0.01, * P < 0.001

Article Snippet: 293FT and NSC34 cell lines from Procell (Wuhan, China) were cultured in DMEM containing 10% fetal bovine serum (FBS) at 37 °C.

Techniques: CCK-8 Assay, In Vitro, Flow Cytometry, Mutagenesis, Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Molecular Medicine

Article Title: The therapeutic potential of Apigenin in amyotrophic lateral sclerosis through ALDH1A2/Nrf2/ARE signaling

doi: 10.1186/s10020-024-00977-7

Figure Lengend Snippet: Effects of ALDH1A2 knockdown and APG treatment on ALS model and OS markers. A - B Validation and selection of the most effective sh-ALDH1A2 to suppress ALDH1A2 expression by qRT-PCR and Western blot analysis. C Flow cytometry to evaluate apoptosis in NSC34 cells treated with APG and sh-ALDH1A2. D Western blot to evaluate the levels of apoptosis-related proteins (Bcl-2, caspase-3, Bax) in NSC34 cells treated with both APG and sh-ALDH1A2. E Flow cytometric analysis of ROS levels in treated NSC34 cells. F - I ELISA for quantification of OS markers (MDA, 4-HNE, SOD, ALDH) in NSC34 cells treated with APG and sh-ALDH1A2. J Western blot analysis of ALDH1A2 protein levels. K - L qRT-PCR and Western blot analysis of Nrf2, HO-1, NQO1 mRNA and protein levels in NSC34 cells treated with APG and sh-ALDH1A2. * P < 0.05, ** P < 0.01, * P < 0.001

Article Snippet: 293FT and NSC34 cell lines from Procell (Wuhan, China) were cultured in DMEM containing 10% fetal bovine serum (FBS) at 37 °C.

Techniques: Knockdown, Biomarker Discovery, Selection, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: The effects of C5-substituted 2,4-diaminoquinazolines on selected transcript expression in spinal muscular atrophy cells

doi: 10.1371/journal.pone.0180657

Figure Lengend Snippet: Clone 11 NSC-34 cells harboring a reporter gene driven by the 3.4-kb SMN2 promoter were treated with 1 μM D156844, D158872, D157161, D157495 or DMSO (n = 4/drug) for 19 hours prior to fluorescent β-lactamase assay analysis. (A) All 4 compounds significantly increased SMN2 -drived BLA activity. The asterisk (*) denotes a statistically significant (p ≤ 0.05) difference between drug- and vehicle-treated cells. Dose-response curves (1 nM– 10 μM) for D156844 (B) , D158872 (C) , D157161 (D) and D157495 (E) . Each compound tested exhibited a dose-dependent increase in SMN2 -drived BLA activity.

Article Snippet: The clone 11 cell line (Vertex Pharmaceuticals, [ ]) was used for the SMN2 promoter assay and the clone 5.3 (Vertex Pharmaceuticals, [ ]) was used for the SMN2 splicing assay.

Techniques: Lactamase Assay, Activity Assay

Journal: PLoS ONE

Article Title: The effects of C5-substituted 2,4-diaminoquinazolines on selected transcript expression in spinal muscular atrophy cells

doi: 10.1371/journal.pone.0180657

Figure Lengend Snippet: EC 50 s of the C5-substituted 2,4-DAQs on SMN2 -drived BLA activity.

Article Snippet: The clone 11 cell line (Vertex Pharmaceuticals, [ ]) was used for the SMN2 promoter assay and the clone 5.3 (Vertex Pharmaceuticals, [ ]) was used for the SMN2 splicing assay.

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: The effects of C5-substituted 2,4-diaminoquinazolines on selected transcript expression in spinal muscular atrophy cells

doi: 10.1371/journal.pone.0180657

Figure Lengend Snippet: (A) Clone 5.3 NSC-34 cells harbor a reporter gene whose expression is linked to inclusion of SMN2 exon 7. These cells were treated with 1 μM D156844, D158872, D157161, D157495 or DMSO (n = 4/drug) for 19 hours prior to fluorescent β-lactamase assay analysis. These compounds tested did not increase the inclusion of SMN2 exon 7 but D157161 and D157495 actually decreased SMN2 exon 7 inclusion. The asterisk (*) denotes a statistically significant (p ≤ 0.05) difference between drug- and vehicle-treated cells. (B) Qualitative analysis of the effects of D156844, D157161, D158872 and D157495 on SMN2 transcripts in type II SMA fibroblasts. GM03813 cells were treated with 1 μM each compound or DMSO (n = 3/compound) for 5 days and then analyzed for changes in the amounts of FL-SMN and SMNΔ7 transcripts by RT-PCR and agarose gel electrophoresis. The amounts of FL-SMN and SMNΔ7 transcripts were also compared against GM03814 samples. COL3A transcripts were also assayed as a loading control for RT-PCR. These compounds tested did not affect the proportion of FL-SMN relative to SMNΔ7 .

Article Snippet: The clone 11 cell line (Vertex Pharmaceuticals, [ ]) was used for the SMN2 promoter assay and the clone 5.3 (Vertex Pharmaceuticals, [ ]) was used for the SMN2 splicing assay.

Techniques: Expressing, Lactamase Assay, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control