Journal:

Article Title: Enrichment for murine keratinocyte stem cells based on cell surface phenotype

doi:

Figure Lengend Snippet: mAb 10G7 recognizes the human transferrin receptor (HTR) CD71. Flow cytometric analysis of CEF + HTR cells showing their reactivity with a known anti-CD71 antibody (B3/25.4) and mAb 10G7. mAb 1D4.5 is an isotype-matched negative control for mAb 10G7.

Article Snippet: Cells were processed for two-color staining for α 6 and CD71 for FACS by incubating with mAb GoH3 (anti-α 6 integrin rat monoclonal; IgG 2a ) at 10 μg/ml (Serotec), and mAb R17 208.2 (a rat anti-mouse CD71 IgM, provided by Bob Hyman, Salk Institute) used as hybridoma supernatant for 1 h, after blocking in 4-ml blocking buffer (4% normal human serum/0.4% BSA/5% FCS in HBSS) for 20 min at 4°C.

Techniques: Negative Control

Journal:

Article Title: Enrichment for murine keratinocyte stem cells based on cell surface phenotype

doi:

Figure Lengend Snippet: Two-color flow cytometric analysis of α6 integrin and CD71 expression on primary murine dorsal keratinocytes. α6 was detected with FITC (x axis, FL1) and CD71 with phycoerythrin (y axis, FL2). Three phenotypically distinct fractions of α6 positive keratinocytes were consistently discernable (n = 25) as indicated: a, α6briCD71bri cells making up the majority of basal keratinocytes; b, α6briCD71dim cells representing a discrete but minor proportion of basal keratinocytes; and c, α6dim cells that appear as a less discrete population. A number of α6 negative nonepithelial cells were also detected.

Article Snippet: Cells were processed for two-color staining for α 6 and CD71 for FACS by incubating with mAb GoH3 (anti-α 6 integrin rat monoclonal; IgG 2a ) at 10 μg/ml (Serotec), and mAb R17 208.2 (a rat anti-mouse CD71 IgM, provided by Bob Hyman, Salk Institute) used as hybridoma supernatant for 1 h, after blocking in 4-ml blocking buffer (4% normal human serum/0.4% BSA/5% FCS in HBSS) for 20 min at 4°C.

Techniques: Expressing

Journal:

Article Title: Enrichment for murine keratinocyte stem cells based on cell surface phenotype

doi:

Figure Lengend Snippet: Cell size and nuclear and cytoplasmic areas of fractionated keratinocytes

Article Snippet: Cells were processed for two-color staining for α 6 and CD71 for FACS by incubating with mAb GoH3 (anti-α 6 integrin rat monoclonal; IgG 2a ) at 10 μg/ml (Serotec), and mAb R17 208.2 (a rat anti-mouse CD71 IgM, provided by Bob Hyman, Salk Institute) used as hybridoma supernatant for 1 h, after blocking in 4-ml blocking buffer (4% normal human serum/0.4% BSA/5% FCS in HBSS) for 20 min at 4°C.

Techniques:

Journal:

Article Title: Enrichment for murine keratinocyte stem cells based on cell surface phenotype

doi:

Figure Lengend Snippet: Distribution of LRCs and PLCs in primary fractionated keratinocytes

Article Snippet: Cells were processed for two-color staining for α 6 and CD71 for FACS by incubating with mAb GoH3 (anti-α 6 integrin rat monoclonal; IgG 2a ) at 10 μg/ml (Serotec), and mAb R17 208.2 (a rat anti-mouse CD71 IgM, provided by Bob Hyman, Salk Institute) used as hybridoma supernatant for 1 h, after blocking in 4-ml blocking buffer (4% normal human serum/0.4% BSA/5% FCS in HBSS) for 20 min at 4°C.

Techniques:

Journal:

Article Title: Enrichment for murine keratinocyte stem cells based on cell surface phenotype

doi:

Figure Lengend Snippet: The bulge region of the hair follicle is CD71dim. (a) Immunofluorescence micrographs of CD71 staining (green) in dorsal skin illustrating several early anagen hair follicles (arrowheads), with bright staining for CD71 at the base of the follicle. Note the longitudinal section through a mid-anagen hair follicle showing relatively low to negative CD71 expression in the bulge region (block arrows), directly below the sebaceous glands (arrow). (b and c) Dual immunofluorescence for CD71 (green) and nuclei stained with propidium iodide (red) illustrating the presence of CD71dim cells in the bulge region (b) and CD71bri cells in the bulb region (c). Dermal papilla is indicated by the star.

Article Snippet: Cells were processed for two-color staining for α 6 and CD71 for FACS by incubating with mAb GoH3 (anti-α 6 integrin rat monoclonal; IgG 2a ) at 10 μg/ml (Serotec), and mAb R17 208.2 (a rat anti-mouse CD71 IgM, provided by Bob Hyman, Salk Institute) used as hybridoma supernatant for 1 h, after blocking in 4-ml blocking buffer (4% normal human serum/0.4% BSA/5% FCS in HBSS) for 20 min at 4°C.

Techniques: Immunofluorescence, Staining, Expressing, Blocking Assay

Journal: Cell reports

Article Title: PI5P4Kα promotes glucose and iron acquisition to support metabolic fitness in pancreatic cancer

doi: 10.1016/j.celrep.2025.116199

Figure Lengend Snippet: (A) FerroOrange staining in the indicated cell lines transduced with sh scr, sh α#1, or sh α#2. Immunofluorescence microscopy images of FerroOrange and DAPI indicating the representative effect of three independent experiments (scale bar, 20 μm) and quantitation of immunofluorescence microscopy images. Data are presented as signal intensity relative to the sh scr condition and are the average of three independent experiments. (B) Transferrin uptake in the indicated cell lines transduced with sh scr, sh α#1, or sh α#2. Data are presented as signal intensity relative to the sh scr condition and are the average of three independent experiments. (C) Surface staining of CD71 quantified using flow cytometry. Data are presented as median intensity relative to the sh scr condition and are the average of three independent experiments. (D) FerroOrange staining in the indicated cell lines transduced with sh scr, sh α#1, or sh α#2 in the presence of FAC. (E) Quantification of relative cell number in the indicated cell lines transduced with sh scr, sh α#1, or sh α#2 in the absence or presence of ferrostatin, ferrostatin with ferric ammonium citrate (FAC), and FAC alone. Data are presented relative to the sh scr condition for each respective treatment and are the average of three independent experiments. (F) Correlation plots between the expression of PIP4K2A from TCGA, with the indicated gene signatures. (G) Kaplan-Meier overall survival curve of patients with PDAC stratified by the expression levels of genes in the indicated gene signatures. (H–J) The MIA PaCa-2 cells were transduced with sh scr, sh α#1, or sh α#2 in the absence or presence of FAC. (H) Glucose consumption. (I) Lactate production. Data are presented relative to the cell number for each respective treatment and are representative of three independent experiments. (J) Immunoblot assessing GLUT1 and PI5P4Kα levels. Tubulin was used as a loading control. Data are representative of three independent experiments. (B–E, H, and I) Data are presented as mean ± SD. Statistical significance was calculated using unpaired two-tailed Student’s t test without (E, H, and I) or with (B–D) Welch’s correction and with pairwise Wilcoxon rank-sum test (F). ns, not significant, * p < 0.05, ** p ≤ 0.001, and *** p ≤ 0.0001.

Article Snippet: Antibodies used were as follows: PI5P4Kα (5527, Cell Signaling), PARP (46D11, Cell Signaling), Glut1 (73015, Cell Signaling), and α-tubulin (T6199, Sigma), p62 (H00008878, Abnova), LC3 (12741, Cell Signaling), p-Histone H3 (9701, Cell Signaling), ASNS (14681–1-AP, Proteintech) and cleaved caspase 3 (9664, Cell Signaling) and PE-conjugated anti-CD71 antibody (130–115-029, Miltenyi Biotec).

Techniques: Staining, Transduction, Immunofluorescence, Microscopy, Quantitation Assay, Flow Cytometry, Expressing, Western Blot, Control, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD71 , REA902 , 50 , 130-115-028 , FITC (PE) , Miltenyi Biotec.

Techniques: Imaging