Journal: Frontiers in Microbiology

Article Title: Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata

doi: 10.3389/fmicb.2015.00843

Figure Lengend Snippet: Strains and plasmids used in this study .

Article Snippet: C. glabrata ATCC 55/ GFP , C. glabrata ATCC 55 (pY13- GFP ) , This study.

Techniques: Mutagenesis

Journal: Frontiers in Microbiology

Article Title: Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata

doi: 10.3389/fmicb.2015.00843

Figure Lengend Snippet: Growth assays in different YNB media. (A) Deletion of CgASG1 has no effect on the utilization of non-fermentative carbon sources in C. glabrata . (B) CgAsg1p plays a role under acid-stress conditions. (C) CgHAL9 serves an important role in cell growth under hypersaline conditions in C. glabrata . (D) CgHal9p plays a role under acid-stress conditions. Logarithmic-phase cells of each C. glabrata strain were adjusted to 2 × 10 7 cells/mL, and then 4 μL of serial tenfold dilutions were spotted onto the corresponding YNB media, as indicated. Pictures were taken after 4 days of growth at 30°C.

Article Snippet: C. glabrata ATCC 55/ GFP , C. glabrata ATCC 55 (pY13- GFP ) , This study.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata

doi: 10.3389/fmicb.2015.00843

Figure Lengend Snippet: Intracellular pH of wild-type, mutant and complemented strains of C. glabrata as below ^a .

Article Snippet: C. glabrata ATCC 55/ GFP , C. glabrata ATCC 55 (pY13- GFP ) , This study.

Techniques: Mutagenesis

Journal: Frontiers in Microbiology

Article Title: Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata

doi: 10.3389/fmicb.2015.00843

Figure Lengend Snippet: Quantitative reverse-transcribed PCR (qRT-PCR) validation of transcriptional profiles in an acidic environment . Logarithmic-phase C. glabrata cells were incubated in YNB or YNB-pH 2.0 media for 2 h. qRT-PCR analyses of the indicated genes was performed as described in the Materials and Methods Section. The means and standard deviations for three independent experiments are shown. C. glabrata strains: wild-type, Cgasg1 Δ and Cghal9 Δ strains. ( * P < 0.05 compared to the corresponding wild-type control, as determined by t -test).

Article Snippet: C. glabrata ATCC 55/ GFP , C. glabrata ATCC 55 (pY13- GFP ) , This study.

Techniques: Reverse Transcription, Quantitative RT-PCR, Biomarker Discovery, Incubation, Control

Journal: Cell

Article Title: A Public BCR Present in a Unique Dual-Receptor-Expressing Lymphocyte from Type 1 Diabetes Patients Encodes a Potent T Cell Autoantigen

doi: 10.1016/j.cell.2019.05.007

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: IRR-MRC-5 iRRADIATED Fibroblast , American Type Cell Culture (ATCC) , Cat#ATCC 55-X.

Techniques: Purification, Virus, Cell Culture, Recombinant, Gel Extraction, Plasmid Preparation, cDNA Synthesis, Protein Quantitation, Irradiation, Cloning, Amplification, Nested PCR, Expressing, Software

Journal: Cancers

Article Title: p62-Induced Cancer-Associated Fibroblast Activation via the Nrf2-ATF6 Pathway Promotes Lung Tumorigenesis

doi: 10.3390/cancers13040864

Figure Lengend Snippet: Post-translational loss of p62 during CAF activation. ( A – F ) Immunofluorescence analysis of p62 expressions in the tumors of 46 stage I/II lung adenocarcinoma patients using a tissue array. αSMA-stained CAFs were distinguished from unstained cancer cells or normal cells. ( A ) The association between p62 expression levels (negative, weak, moderate, and strong) and cancer stage. G2 and G3: grading states. ( B ) The correlation between p62 and αSMA expressions. ( C , D ) Representative images showing strong p62 expressions in CAF-adjacent tumors. CAFs were surrounded with yellow line and cancer cells were indicated with white arrows. Scale bars: ( C ) 200 μm, ( D ) 20 μm. ( E ) Quantification of intensity signal for p62. Error bars, SD. ( F ) The association between p62 expressions and the distance from CAF (<200 μm or >200 μm). Error bars, SD. ( G ) A schematic representation of CAF differentiation from normal fibroblasts by co-culture with GFP-tagged cancer cells. ( H ) Representative images of immunofluorescence staining revealing p62 expression levels (red) in A549-GFP cells (green, indicated with arrows) after the co-culture with MRC5 cells. Scale bars: 20 μm. ( I , J ) Immunofluorescence analysis of p62 expressions in the CAFs of 46 stage I/II lung adenocarcinoma patients using a tissue array. αSMA-stained CAFs were distinguished from unstained cancer cells or normal cells. ( I ) Representative images showing p62 expressions (green) in CAFs. The nuclei were stained with DAPI (blue). Scale bars: 20 μm. ( J ) Quantification of intensity signal for p62. Error bars, SD. ( K ) Analysis of P62 mRNA expressions in the CAFs of 9 lung adenocarcinoma patients (6 for stage I/II and 3 for stage III). The association between P62 expression levels and cancer stages. For statistical analysis, two-tailed Mann-Whitney U test was used. Error bars, SD. ( L ) The mRNA expressions of P62 in MRC5 cells after hypoxia (1% O 2 , 24 h) were analyzed by Real-time RT PCR. GAPDH was used as a reference gene in the analysis. Error bars, SD ( n = 3). ( M ) The protein expressions of p62 in MRC5 cells after hypoxia (1% O 2 , 24 h) were assessed with Western blot analysis. Full-length blots are presented in . ( N ) Quantification of ( M ). Error bars, SD ( n = 3). ( O ) Representative images of immunofluorescence staining revealing p62 expressions (red) in MRC5 cells (surrounded with yellow line) after the co-culture with A549-GFP cells (green, indicated with white arrows) for 4 days. Scale bars: 20 μm. ( P ) Quantification of intensity signal for p62 in ( O ) Error bars, SD. (* p < 0.05, ** p < 0.001, p -values between depicted groups.).

Article Snippet: Human fetal lung fibroblasts (MRC5, Cat#10171), human lung adenocarcinoma cells (A549, Cat#CCL-185), and GFP-labeled A549 cells (A549-GFP, Cat#MBS168483) were purchased from the Korean Cell Line Bank (Seoul, Korea), the American Type Culture Collection (Manassas, VA, USA), and MyBioSource (San Diego, CA, USA), respectively. p62 +/+ and p62 −/− mouse embryonic fibroblasts (MEFs) were obtained from Keiji Tanaka’s laboratory (Tokyo Metropolitan Research Institute, Tokyo, Japan) with Tetsuro Ishii’s permission.

Techniques: Activation Assay, Immunofluorescence, Staining, Expressing, Co-Culture Assay, Two Tailed Test, MANN-WHITNEY, Quantitative RT-PCR, Western Blot

Journal: Cancers

Article Title: Oncogenic and Stemness Signatures of the High-Risk HCMV Strains in Breast Cancer Progression

doi: 10.3390/cancers14174271

Figure Lengend Snippet: Replication of B544 and B693 strains in MRC5 cultures, and the appearance of morphologically distinct cells following the infection of HMECs with these high-risk strains. ( A ) Time-course of the viral titer in the supernatant of MRC5 infected with the strains HCMV-B544 and HCMV-B693, as measured by IE1-qPCR. ( B ) Confocal microscopic images of HCMV-IE1 and pp65 staining in HMECs infected with HCMV-B544 and HCMV-B693 (day 1 post-infection). Uninfected HMECs were used as controls. Nuclei were counterstained with DAPI; magnification ×63, scale bar 10 μm. ( C ) HMECs time-course infection with HCMV-B544 and HCMV-B693 strains (MOI = 1). Magnification ×100, scale bar 100 μm. Uninfected HMECs were used as a control. ( D ) Presence of giant cells with blastomere-like morphology (1 and 6), mesenchymal cells (4 and 7), lipid droplet-packed cells (3, 8, and 9), cells displaying multiple nuclei (2) as well as cell budding (4, 5, and 6), and cells with filopodia protrusions (9) in CTH-B544 and CTH-B693 cells. The inverted light microscope scale bar represents 100 µm; magnification ×200.

Article Snippet: HMECs (A10565, Life Technologies, Carlsbad, CA, USA), CTH cells, MDA-MB231 as well as MCF7 (Institut Hiscia, Arlesheim, Switzerland), and MRC5 (RD-Biotech, Besançon, France) were cultured as previously described [ ].

Techniques: Infection, Staining, Light Microscopy

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Pulmonary activation of vitamin D 3 and preventive effect against interstitial pneumonia

doi: 10.3164/jcbn.19-48

Figure Lengend Snippet: Expression of the vitamin D receptor and vitamin D-metabolizing enzymes in human fibroblasts, and VD 3 -dependent suppression of the induction of IL-1β gene expression in pulmonary fibroblasts by bleomycin. (A) To assess gene expression in MRC-5 and MRC-5 SV1 TG1 cells, RT-PCR was performed using specific primers for the vitamin D receptor (VDR) or vitamin D-metabolizing enzymes, including CYP27A1 (27A1), CYP2R1 (2R1), and CYP27B1 (27B1). (B) MRC-5 SV1 TG1 cells were treated with 25 µg/ml bleomycin for 48 h and RT-PCR was performed using specific primers for αSMA, IL-1β, and GAPDH. (C, D) Band intensities of the PCR products were converted to numerical data by image analysis software, and the data for αSMA and IL-1β were normalized by the GAPDH value.

Article Snippet: A normal human fetal pulmonary fibroblast cell line (MRC-5) and an immortalized cell line derived from MRC-5 (MRC-5 SV1 TG1) (KAC Co., Ltd., Kyoto, Japan) were maintained in α-minimal essential medium (α-MEM) containing 10% fetal calf serum (FCS) and antibiotics (Life Technologies Japan Ltd., Tokyo, Japan) at 37°C under 5% CO 2 .

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Software

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Pulmonary activation of vitamin D 3 and preventive effect against interstitial pneumonia

doi: 10.3164/jcbn.19-48

Figure Lengend Snippet: Pretreatment of pulmonary fibroblasts with VD 3 suppresses bleomycin-induced expression of pro-fibrotic genes, fibrosis markers, and inflammatory markers. (A) MRC-5 SV1 TG1 cells underwent pretreatment with VD 3 or 1,25(OH) 2 D 3 overnight and then were incubated with 25 µg/ml bleomycin for 24 h, after which RT-PCR was performed using specific primers for αSMA, COLA2, SPP1, IL-1β, TGF-β1, and β-actin (ACTB). (B–D) Band intensities of the PCR products were converted to numerical data by image analysis software, and the data for SPP1, IL-1β, and TGF-β1 were normalized by the β-actin value.

Article Snippet: A normal human fetal pulmonary fibroblast cell line (MRC-5) and an immortalized cell line derived from MRC-5 (MRC-5 SV1 TG1) (KAC Co., Ltd., Kyoto, Japan) were maintained in α-minimal essential medium (α-MEM) containing 10% fetal calf serum (FCS) and antibiotics (Life Technologies Japan Ltd., Tokyo, Japan) at 37°C under 5% CO 2 .

Techniques: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Software

Journal: BMC Pulmonary Medicine

Article Title: The effect of the mechanodynamic lung environment on fibroblast phenotype via the Flexcell

doi: 10.1186/s12890-024-03167-7

Figure Lengend Snippet: Summary of all Flexcell studies assessing the effects of the mechanical lungs on fibroblast phenotype and function

Article Snippet: MRC-5 lung fibroblasts 2D culture on BioFlex plates with LPS exposure , 5%, 10%, 15%, 20% , 0.1 Hz , 5% and 10% amplitudes increased MRC-5 proliferation, while 15% and 20% decreased proliferation rates , [ ] .

Techniques: Knock-Out, Comparison, Concentration Assay, Expressing, Phospho-proteomics, Control