Journal: PLoS ONE

Article Title: The Anti-Tumor Histone Deacetylase Inhibitor SAHA and the Natural Flavonoid Curcumin Exhibit Synergistic Neuroprotection against Amyloid-Beta Toxicity

doi: 10.1371/journal.pone.0085570

Figure Lengend Snippet: A. PC12 cells were treated with different concentrations of Aβ 25–35 (n = 5). *, p <0.05; ***, p <0.001 vs Ctrl. B. PC12 cells were treated with different concentrations of SAHA (n = 5). *, p <0.05; ***, p <0.001 vs Ctrl. C. PC12 cells were treated with different concentrations of curcumin (n = 5). *, p <0.05; ***, p <0.001 vs Ctrl. D. MTT assay was performed to detect cell viability after treating with SAHA and curcumin against Aβ 25–35 -induced cytotoxicity in PC12 cells (n = 5). ***, p <0.001 vs Ctrl; **, p <0.01 vs Aβ 25–35 .

Article Snippet: Rat pheochromocytoma PC12 cells were originally obtained from Chinapeptides Co., Ltd. PC12 neuron cells were cultured in DMEM-F12 containing 7% FBS, 1% penicillin, and 1% streptomycin at 37°C.

Techniques: MTT Assay

Journal: PLoS ONE

Article Title: The Anti-Tumor Histone Deacetylase Inhibitor SAHA and the Natural Flavonoid Curcumin Exhibit Synergistic Neuroprotection against Amyloid-Beta Toxicity

doi: 10.1371/journal.pone.0085570

Figure Lengend Snippet: A. Apoptotic cells were detected by TUNEL assay. Cells were stained with TUNEL positive nuclei (green) and nuclei of PC12 cells (blue). B. The percentage of TUNEL positive cells was determined (n = 5). ***, p <0.001 vs Ctrl; **, p <0.01 vs Aβ 25–35 . Ctrl: the control group. C. Western blot of cleaved caspase 3 (n = 3). ***, p <0.001 vs Ctrl; **, p <0.01 vs Aβ 25–35 . Ctrl: the control group; CASP3: caspase 3.

Article Snippet: Rat pheochromocytoma PC12 cells were originally obtained from Chinapeptides Co., Ltd. PC12 neuron cells were cultured in DMEM-F12 containing 7% FBS, 1% penicillin, and 1% streptomycin at 37°C.

Techniques: TUNEL Assay, Staining, Control, Western Blot

Journal: PLoS ONE

Article Title: The Anti-Tumor Histone Deacetylase Inhibitor SAHA and the Natural Flavonoid Curcumin Exhibit Synergistic Neuroprotection against Amyloid-Beta Toxicity

doi: 10.1371/journal.pone.0085570

Figure Lengend Snippet: A. Representative western blot of the p -Akt (Ser 473) protein expression with SAHA treatment (n = 3). **, p <0.01 vs Ctrl. B. Representative western blot of the p -Akt (Ser 473) protein expression with curcumin treatment (n = 3). **, p <0.01 vs Ctrl. C. Representative western blot of the p -Akt (Ser 473) protein expression with co-treatment of SAHA and curcumin (n = 3) to the PC12 cells (n = 3). *, p <0.05 vs Ctrl; **, p <0.01 vs Aβ 25–35 ; Ctrl: the control group.

Article Snippet: Rat pheochromocytoma PC12 cells were originally obtained from Chinapeptides Co., Ltd. PC12 neuron cells were cultured in DMEM-F12 containing 7% FBS, 1% penicillin, and 1% streptomycin at 37°C.

Techniques: Western Blot, Expressing, Control

Journal: Neural Regeneration Research

Article Title: Srgap2 suppression ameliorates retinal ganglion cell degeneration in mice

doi: 10.4103/1673-5374.369122

Figure Lengend Snippet: Downstream signaling pathways after inhibition of Srgap2 . (A) Detection of SRGAP2 (green, Alexa Fluor 488) expression in PC12 cells by immunofluorescence staining. n = 3 independent cell culture preparations. Scale bars: 50 μm. (B) SRGAP2 protein level was decreased after siRNA transfection and Rac1 and Cdc42 were increased after inhibiting Srgap2. (C) Quantification of B. n = 3 independent cell culture preparations. (D) mTOR signaling pathway was activated after the downregulation of Srgap2. (E) Quantification of D. n = 3 independent cell culture preparations. (F, G) ONC induced activation of the mTOR signaling pathway 7 days after ONC. Srgap2 +/– mice showed increased mTOR activation compared with WT. n = 3 independent animals in each group. Data (normalized by WT-ctrl group) are presented as mean ± SEM. ** P < 0.01, *** P < 0.001, **** P < 0.0001 (two-tailed Student’s t -test). Cdc42: Cell division cycle 42; Ctrl: control; DAPI: 4′,6-diamidino-2-phenylindole; mTOR: mammalian target of rapamycin; ONC: optic nerve crush; Rac1: Ras-related C3 botulinum toxin substrate 1; siRNA: small interfering RNA; Srgap2: Slit-Robo GTPase activating protein 2; WT: wild type.

Article Snippet: PC12, one of the most commonly used cell lines in neuroscience research (You et al., 2022), was purchased from CHI Scientific (Jiangyin, China; RRID: CVCL_0481).

Techniques: Protein-Protein interactions, Inhibition, Expressing, Immunofluorescence, Staining, Cell Culture, Transfection, Activation Assay, Two Tailed Test, Control, Small Interfering RNA

Journal: Cell Death & Disease

Article Title: MiR-127-3p targeting CISD1 regulates autophagy in hypoxic–ischemic cortex

doi: 10.1038/s41419-021-03541-x

Figure Lengend Snippet: A Results of miRNA sequencing analysis showed that the expression of miRNAs in sham and HI have obvious difference ( n = 2/group). B The qRT-PCR validation of differential miRNAs in normal group rats and HI group rats at 12 h after HI injury. Each bar was the mean ± s.d. relative to −log2 (** P < 0.01 with Student’s t -test, n = 6). C Relative expression of miRNA-127-3p in heart, liver, brain, and kidney of rats. MiRNA-127-3p was selectively highly expressed in the brain (** P < 0.01 with one-way ANOVA, n = 6). D qRT-PCR analysis of miR-127-3p expression at 12, 48, and 96 h. MiRNA-127-3p decreased at 12 h after HI, while it increased at 48 and 96 h after HI injury (** P < 0.01 with one-way ANOVA, n = 6). E The relative expression of miR-127-3p in OGD of PC12, SY5Y, and neurons was significantly decreased in vitro (** P < 0.01 with Student’s t -test, n = 6). HI hypoxic ischemic.

Article Snippet: PC12 cell line (GCC-KI0009RT) was obtained from Shanghai Genechem Co., LTD and SY5Y cell line (BFN60700126) was obtained from BLUEFBIO.

Techniques: Sequencing, Expressing, Quantitative RT-PCR, Biomarker Discovery, In Vitro

Journal: PLoS ONE

Article Title: NGF and proNGF Regulate Functionally Distinct mRNAs in PC12 Cells: An Early Gene Expression Profiling

doi: 10.1371/journal.pone.0020839

Figure Lengend Snippet: Panel A: Schematic representation of the short form of proNGF . The arrows mark the cleavage sites for furin, the double headed arrows represent the C-terminal processing site (post translational modification) and hexagons the potential N-glycosylation sites. In red, the di-basic amino acids that are important in the processing of the protein. In green, the consensus site for the cleavage by furin. In the present study, we obtained furin resistant mutants in this site (mutant proNGF-KR: RS KR to RS AA ). Panel B: Western blot to test the stability of the proNGF samples in PC12 cells . PC12 cells were plated and treated for 1 h or 4 h with 20 ng/mL of proNGF-WT or proNGF- KR or 10 ng/mL of NGF. Then, the medium was taken and 1 µg of recombinant proNGF or proNGF-KR or NGF was spiked into 50 µL of conditioned medium. Spiking controls in fresh medium and PBS were also carried out. The spiked medium was incubated at 37°C for 1 h or 4 h. The red arrow marks the band corresponding to the full length proNGF, the blue arrow marks the band corresponding to mature NGF. In the figure, WT stands for proNGF-WT and KR stands for proNGF-KR. Panel C: Densitometric analysis performed on the Western Blot of the spiking experiments representing the percentage of proNGF proteolysis . The bands corresponding to proNGF and NGF in the Western blot challenged with the anti-NGF antibody were quantified. The resulting intensities, normalized against the areas of the bands, were reported in the histogram. For each lane, corresponding to the different proNGF-WT or -KR treatments, the band intensities of proNGF and NGF, derived from proNGF proteolysis, were measured and the sum of the two bands intensities was assigned to a value of 100%. Among this total intensity, the intensity of the bands corresponding to proNGF and mature NGF was evaluated and expressed as %. The histogram is the result of the average of four independent experiments.

Article Snippet: Rat pheochromocytoma PC12 cells (PC12 SB subclone, kindly provided by Maurizia Caruso, Consiglio Nazionale delle Ricerche, INMM, Rome, Italy) were maintained with RPMI 1640 Medium (Invitrogen) and grown as monolayer cultures on Falcon dishes, supplemented with 10% Horse Serum (Invitrogen) and 5% Foetal Calf Serum (Invitrogen), in a humidified atmosphere at 37°C and 5% CO 2 .

Techniques: Modification, Glycoproteomics, Mutagenesis, Western Blot, Recombinant, Incubation, Derivative Assay

Journal: PLoS ONE

Article Title: NGF and proNGF Regulate Functionally Distinct mRNAs in PC12 Cells: An Early Gene Expression Profiling

doi: 10.1371/journal.pone.0020839

Figure Lengend Snippet: Phase contrast pictures (upper two rows, 20× magnification) of PC12 cells in culture and confocal images of triple immunofluorescence of PC12 cells (last four rows) respectively for betaIII-tubulin (green), actin (red) and DNA (blue) (63× magnification) and immunofluorescence for actin cytoskeleton (100× magnification). After 1 h or 4 h cells were fixed and stained with anti-betaIII tubulin antibody, Alexa 594 phalloidin to visualize filamentous actin and DAPI for nuclear staining. - first column: control cells with no addition. - second column: 10 ng/mL of NGF. - third column: 20 ng/mL of proNGF WT. - fourth column: 20 ng/mL of proNGF KR.

Article Snippet: Rat pheochromocytoma PC12 cells (PC12 SB subclone, kindly provided by Maurizia Caruso, Consiglio Nazionale delle Ricerche, INMM, Rome, Italy) were maintained with RPMI 1640 Medium (Invitrogen) and grown as monolayer cultures on Falcon dishes, supplemented with 10% Horse Serum (Invitrogen) and 5% Foetal Calf Serum (Invitrogen), in a humidified atmosphere at 37°C and 5% CO 2 .

Techniques: Immunofluorescence, Staining, Control

Journal: Acta Pharmacologica Sinica

Article Title: hERG channel function: beyond long QT

doi: 10.1038/aps.2013.6

Figure Lengend Snippet: hERG function and pathology classified by tissue type.

Article Snippet: Adrenal gland , Primary, rat PC12 , Unigene EST , Current inhibition, RNAse protection (cDNA) , Epinephrine release , Adenoma , , , , .

Techniques: Expressing, Functional Assay, Inhibition, Western Blot, Migration, Immunohistochemistry