Journal: Yonsei Medical Journal

Article Title: Changes in Inward Rectifier K + Channels in Hepatic Stellate Cells During Primary Culture

doi: 10.3349/ymj.2008.49.3.459

Figure Lengend Snippet: Relative inward rectifier K + channel and SUR gene expression in HSC. (A) The inward rectifier K + channel α-subunit gene expression in HSC was measured using real-time RT-PCR. The bar graphs show the relative gene expression for each inward rectifier K + channel subfamily (K ir 1.1, K ir 2.1 - K ir 2.4, K ir 3.1 - K ir 3.4, K ir 4.1 - K ir 4.2, K ir 5.1, K ir 6.1 - K ir 6.2, and K ir 7.1). (B) SUR gene expression was measured. The bar graphs show the relative gene expression for SUR1, SUR2A, and SUR2B. The HSC were used at 1 day, 1 week, 2 weeks, and 3 weeks of culture. The expression levels were normalized to GAPDH and calibrated by K ir 2.1 expression in the HSC cultured for 1 day. The data are shown as the mean ± SEM (n = 3). (C) K ir 2.1 and K ir 6.1 protein expression were measured by Western blotting. There was a band (55 kD) in the anti-K ir 2.1 membrane (Upper) as well as the anti-K ir 6.1 membrane (Lower). Both membranes were re-probed with anti-GAPDH antibody as shown. The data is representative of three independent experiments.

Article Snippet: The anti-K ir 2.1 and anti-K ir 6.1 antibodies were obtained from Alomone Labs (Jerusalem, Israel), the anti-GAPDH antibody was acquired from Novus Biologicals (Littleton, CO, USA), and Western Lightning was supplied by PerkinElmer Life Sciences (Gaithersburg, MD, USA).

Techniques: Gene Expression, Quantitative RT-PCR, Expressing, Cell Culture, Western Blot, Membrane

Journal: Cell Reports Methods

Article Title: Development of an HIV reporter virus that identifies latently infected CD4 + T cells

doi: 10.1016/j.crmeth.2022.100238

Figure Lengend Snippet:

Article Snippet: Anti-human CD27 (REA499) 155Gd, coupled in house , Miltenyi Biotec , Cat#: 130122295, RRID: AB_2801876.

Techniques: Virus, Plasmid Preparation, Recombinant, Cell Isolation, Staining, Software

Journal: American Journal of Cancer Research

Article Title: Impact of BMI1 expression on the apoptotic effect of paclitaxel in colorectal cancer

doi:

Figure Lengend Snippet: BMI1 knockdown enhances the cytotoxicity of paclitaxel in CRC cells. A. The level of BMI1 protein was determined using western blot analysis. B. Cytotoxicity of indicated protein kinase inhibitor to HT-29 cells. Cells were treated with protein kinase inhibitor (1 μM PKC412; 10 nM paclitaxel; 10 μM VX-702; 10 μM Imatinib) as indicated for 72 hours, and cell viability was determined via the MTT assay. The experiments were repeated three times. Between-group comparisons were performed using one-way ANOVA (*, P<0.05). C. Morphology of apoptosis was visualized under a light microscope. White arrowheads in the pictures indicate the nuclei of apoptotic cells.

Article Snippet: Anti-BMI1 was purchased from Bethyl Laboratories, Inc. Anti-cleaved caspases-3, cleaved caspase-8, Bcl-2, tubulin, and actin were purchased from Genetex (San Antonio, TX, USA).

Techniques: Western Blot, MTT Assay, Light Microscopy

Journal: American Journal of Cancer Research

Article Title: Impact of BMI1 expression on the apoptotic effect of paclitaxel in colorectal cancer

doi:

Figure Lengend Snippet: Paclitaxel has stronger effect on apoptotic signaling pathways in BMI1-deficient cells. A. Protein levels that cleaved caspase-3 and BMI1 were determined in cells treated with indicated inhibitors using western blot analysis. B. The percentage of apoptotic cells treated with paclitaxel was detected via dual staining with annexin V-FITC and PI in HT-29 cell lines. HT-29 BMI1-deficient (sh-BMI1) cell lines were analyzed via flow cytometry. The experiments were repeated three times. Between-group comparisons were performed using one-way ANOVA (*, P<0.05). C. The percentage of apoptotic cells treated with paclitaxel combined with caspase-8 inhibitor (Z-IETD-FMK) or pan-caspase inhibitor (zVAD-fmk) was detected via dual staining with Annexin V-FITC and PI in HT-29, and HT-29 BMI1-deficient (sh-BMI1) cell lines were analyzed via flow cytometry. The experiments were repeated three times. Between-group comparisons were performed using one-way ANOVA (*, P<0.05).

Article Snippet: Anti-BMI1 was purchased from Bethyl Laboratories, Inc. Anti-cleaved caspases-3, cleaved caspase-8, Bcl-2, tubulin, and actin were purchased from Genetex (San Antonio, TX, USA).

Techniques: Western Blot, Staining, Flow Cytometry

Journal: American Journal of Cancer Research

Article Title: Impact of BMI1 expression on the apoptotic effect of paclitaxel in colorectal cancer

doi:

Figure Lengend Snippet: BMI1 deficiency induces activation of GSK3beta and JNK in modulating the phosphorylation of MCL-1. A. Protein levels of Bak, Bad, Bax, BID, and BMI1 in cells treated with paclitaxel, determined using western blot analysis. B. Protein levels of MCL-1, BCL-2, and BMI1 in cells treated with paclitaxel, determined using western blot analysis. C. Expression of BCL-2 and MCL-1 mRNA in HT-29-sh-Luci, HT-29-sh-BMI1#1 cells, and HT-29-sh-sh-BMI1#2 cells determined using real-time qPCR. Columns represent the mean results from PCR assays performed in triplicate and normalized to GAPDH (*, P<0.05). D. Densitometry was utilized to quantify MCL-1 protein levels after normalization with Actin to obtain the percentage of MCL-2 degradation (mean ± SD; n=3). Error bars indicate SD. E. Protein levels in HT-29-sh-Luci, HT-29-sh-BMI1#1 cells, and HT-29-sh-sh-BMI1#4 cells analyzed via western blot analysis.

Article Snippet: Anti-BMI1 was purchased from Bethyl Laboratories, Inc. Anti-cleaved caspases-3, cleaved caspase-8, Bcl-2, tubulin, and actin were purchased from Genetex (San Antonio, TX, USA).

Techniques: Activation Assay, Western Blot, Expressing

Journal: American Journal of Cancer Research

Article Title: Impact of BMI1 expression on the apoptotic effect of paclitaxel in colorectal cancer

doi:

Figure Lengend Snippet: Bmi1 deficiency combined with paclitaxel retards tumor growth in vivo. A. Paclitaxel treatment in BMI1-deficient cells leads to the reduction of tumor growth. Representative images of HT-29-sh-Luci and HT-29-sh-BMI in SCID mice. B. Quantitative analysis of tumor size after injection in designated cells at 30 days. Statistical comparisons were conducted using one-way ANOVA (*, P<0.05). C. Representative staining for Ki-67, γ-H2AX, MCL-1, and p-GSK3beta in tumors with mock and paclitaxel treatment groups. Original magnification: × 40, scale bar: 10 μm.

Article Snippet: Anti-BMI1 was purchased from Bethyl Laboratories, Inc. Anti-cleaved caspases-3, cleaved caspase-8, Bcl-2, tubulin, and actin were purchased from Genetex (San Antonio, TX, USA).

Techniques: In Vivo, Injection, Staining

Journal: The Journal of Neuroscience

Article Title: Regulation of Myelination by Exosome Associated Retinoic Acid Release from NG2-Positive Cells

doi: 10.1523/jneurosci.2922-18.2019

Figure Lengend Snippet: Figure 7. Raldh2 activity is required for differentiation of NG2+cells into myelinating 795

Article Snippet: 319 Antibodies used were: mouse monoclonal anti-βIII tubulin (Promega, 1:1000, Cat# G7121, 320 RRID:AB_430874); chicken polyclonal anti- GFAP (Abcam, 1:300, Cat# ab4674, 321 RRID:AB_304558); rabbit polyclonal anti- GFAP (DAKO, 1:2500, ); mouse monoclonal 322 anti- GFAP (Sigma, 1:100, Cat# Z0334, RRID:AB_10013382); rabbit polyclonal anti- 323 14 RAR (Santa Cruz Biotechnology, Inc, 1:100, Cat# sc-552, RRID:AB_2175379); rabbit 324 polyclonal anti-NG2 (Millipore, 1:100, Cat# 05-710, RRID:AB_309925); goat polyclonal 325 anti- aldehyde dehydrogenase1A2 (Raldh2) (Santa Cruz Biotechnology, Inc, 1:100, Cat# sc-326 22591, RRID:AB_2224036); mouse monoclonal anti-CSPG (CS-56) (Sigma, 1:100, Cat# 327 C8035, RRID:AB_476879), goat polyclonal anti-decorin (R&D Systems, 1:20, for 328 immunochemistry and 5 g/ml as a neutralizing antibody, Cat# AF143, RRID:AB_354790), 329 rabbit polyclonal anti-decorin (Abcam, 1:50, Cat# ab175404), rat monoclonal anti-CD44 330 (eBioscience, 1:150, Cat# 14-0441-86, RRID:AB_467248), mouse monoclonal anti- myelin 331 associated protein (MAG) (Abcam, 1:2000, Cat# ab89780, RRID:AB_2042411); goat 332 polyclonal anti-Olig1 (Abcam, 1:1000, Cat# ab68105, RRID:AB_1142042); rabbit 333 polyclonal anti-myelin proteolipid protein (PLP) (Abcam, 1:1000, Cat# ab105784, 334 RRID:AB_10973392); goat polyclonal anti- RARα (Abcam, 1:100, Cat# ab28767, 335 RRID:AB_777684); rabbit polyclonal anti-oligodendrocyte specific protein (OSP) (Abcam, 336 1:2,000, Cat# ab53041, RRID:AB_2276205); rabbit polyclonal anti-retinoic acid (RA) 337 (Abnova, 1:300, Cat# PAB15482, RRID:AB_10759405); rabbit monoclonal anti-EGFR 338 (Abcam, 1:500, Cat# ab52894, RRID:AB_869579); rabbit polyclonal anti-AIP1/Alix (1:1000 339 for western blotting, Cat# ABC40, RRID:AB_10806218).

Techniques: Activity Assay