Journal: Scientific Reports

Article Title: Choice of lipid supplementation for in vitro erythroid cell culture impacts reticulocyte yield and characteristics

doi: 10.1038/s41598-026-37229-z

Figure Lengend Snippet: qPCR shows cholesterol biosynthesis is significantly increased in plasma cultured cells. ( A ) Main steps in cholesterol and fatty acid biosynthesis. Cholesterol biosynthesis begins with acetyl-CoA and proceeds via HMG-CoA reductase (HMGCR), forming mevalonate and downstream intermediates (mevalonate-PP, isopentenyl-PP, and squalene). Squalene epoxidase (SQLE) converts squalene to 2,3(S)-oxidosqualene, leading to lanosterol and cholesterol synthesis. Fatty acid biosynthesis also originates from acetyl-CoA, with acetyl-CoA carboxylase (ACC) producing malonyl-CoA, followed by fatty acid synthase (FASN) generating palmitate and complex fatty acids. SREBP1/2 transcription factors regulate these pathways, with SREBP1 driving fatty acid synthesis and SREBP2 controlling cholesterol biosynthesis. The LDL receptor (LDLR) mediates cholesterol uptake, maintaining lipid homeostasis. Made using Biorender. ( B ) mRNA levels of HMGCR (HMG-CoA reductase), SQLE (Squalene Epoxidase), LDLR (Low density lipoprotein receptor), SREBP1 and SREBP2 (Sterol regulatory element-binding protein 1 or 2) relative to GAPDH on day 8 of differentiation after CD34 + isolation. A parametric Brown-Forsythe and Welch test was performed to test for differences between groups. p < 0.05 was considered statistically significant ( n = 3, N = 3). ( C ) Example waterfall plots of cells the same donor on day 8 of erythroid differentiation, labelled against Band3 and CD49d, grown in medium containing AB serum, Plasma, or Plasma supplemented with CRL. Corresponding percentage of each stage (I-Proerythoblast, II-Basophilic, III-Polychromatic, IV-Orthochromatic, and V-Reticulocyte) is indicated next to the corresponding gate. D Shows percentage of erythroid cell types present on day 8 for each condition studied, quantified as displayed in panel above ( n = 3). Error bars represent the standard deviation.

Article Snippet: CD49d was detected using a FITC-conjugated anti-human CD49d antibody (clone MZ18-24A9), a mouse IgG2b from Miltenyi Biotech.

Techniques: Clinical Proteomics, Cell Culture, Binding Assay, Isolation, Standard Deviation

Journal: REPRODUCTION

Article Title: Involvement of VCAM1 in the bovine conceptus adhesion to the uterine endometrium

doi: 10.1530/rep-13-0655

Figure Lengend Snippet: Figure 5 Expression of VCAM1 receptor, integrin a 4 (ITGA4), by bovine conceptuses. (A) Changes in ITGA4 mRNAs in days 17, 20, and 22 conceptuses (P17, P20, and P22 respectively). Note that ITGA4 transcripts were found minimal at the uterine epithelium and stroma. Values represent meanGS.E.M. from three independent samples with duplicates within a day of conceptus collection. **Statistically significant differences in mRNA levels (P!0.01). (B) Immunohisto- chemical analysis of ITGA4 (a, b, c, and d) or VCAM1 (e, f, g, and h) expression in the bovine uterus obtained from day 22 pregnant animals. Tissue sections (10 mm) from day 22 uteri were immunostained for ITGA4 using an anti-ITGA4 antibody (a and c) or normal mouse IgG as a negative control at low (b) and higher magnification (d). (c) Observation of boxed area in B-a at a higher magnification. Tissue sections were immunostained for VCAM1 using an anti-VCAM1 antibody (e and g), or normal rabbit IgG as a negative control at low (f) and higher magnification (h). (g) Observation of boxed area in B-e at a higher magnification. Epi, endometrial luminal epithelium; St, endometrial stroma; Tr, trophoblast. Black scale barZ200 mm and white scale barZ40 mm.

Article Snippet: After 30 min of incubation with 10% normal goat serum, the sections were incubated at 4 8C overnight with a rabbit anti-human VCAM1 polyclonal antibody (1:100 dilution, 0.5 mg/ml, ab106777, Abcam, Cambridge, MA, USA), a mouse anti-human CD49d (ITGA4) antibody (1:100 dilution, 1 mg/ml, MCA697GA, AbD Serotec, Hercules, CA, USA), or normal mouse IgG (1:40 dilution, 0.4 mg/ml, sc-2025, Santa Cruz Biotechnology, Inc.) or normal rabbit IgG (1:80 dilution, 0.4 mg/ml, sc-2027, Santa Cruz Biotechnology, Inc.) as a Reproduction (2014) 148 119–127 A negative control.

Techniques: Expressing, Negative Control

Journal: eLife

Article Title: Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

doi: 10.7554/eLife.60933

Figure Lengend Snippet:

Article Snippet: Antibody , CD49d (α4) (mouse monoclonal) , Fluidigm , Cat#3141004B , (1 μg/100 μL).

Techniques:

Journal: eLife

Article Title: Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

doi: 10.7554/eLife.60933

Figure Lengend Snippet:

Article Snippet: Antibody , CD49d (α4) (mouse monoclonal) , Fluidigm , Cat#3141004B , (1 μg/100 μL).

Techniques:

Journal: iScience

Article Title: hESC- and hiPSC-derived Schwann cells are molecularly comparable and functionally equivalent

doi: 10.1016/j.isci.2024.109855

Figure Lengend Snippet: hESC and hiPSC cells are efficiently differentiated into Schwann cells (A) Diagram of the 21-day Schwann cell differentiation protocol. (B) Representative cells count graphs of CD49d FACS sorted hESC- and hiPSC-derived Schwann cells. (C) Representative Schwann cell differentiation efficiencies for each hESC and hiPSC line ( n = 1 each). (D) Representative images of passage two CD49d-positive hESC- and hiPSC-derived Schwann cells processed for immunocytochemistry with an anti-S100B antibody (red), Phalloidin to label F-actin (green) and DAPI (blue). Scale bar, 50μm.

Article Snippet: The remaining cells were transferred to a 1.5ml tube, 10μl PE-conjugated anti-CD49d antibody (R&D Systems, FAB1354P) was added and the cells were rotated in the dark at 4°C for 30 minutes.

Techniques: Cell Differentiation, Derivative Assay, Immunocytochemistry

Journal: iScience

Article Title: hESC- and hiPSC-derived Schwann cells are molecularly comparable and functionally equivalent

doi: 10.1016/j.isci.2024.109855

Figure Lengend Snippet:

Article Snippet: The remaining cells were transferred to a 1.5ml tube, 10μl PE-conjugated anti-CD49d antibody (R&D Systems, FAB1354P) was added and the cells were rotated in the dark at 4°C for 30 minutes.

Techniques: Produced, Recombinant, Knock-Out, Electron Microscopy, Staining, Membrane, Gene Expression, Ab Array, Software, Sequencing, Sterility

Journal: The Journal of Experimental Medicine

Article Title: Functional and clinical relevance of VLA-4 (CD49d/CD29) in ibrutinib-treated chronic lymphocytic leukemia

doi: 10.1084/jem.20171288

Figure Lengend Snippet: Triggering of the BCR by anti-IgM induces VLA-4 activation and increases cell adhesion and VLA-4 clustering in CLL cells also upon ibrutinib treatment in vitro. (A) Calcium response to anti-IgM stimulation in PBMCs from 30 CLL cases. The pseudocolored dot plot on the right shows an example of BCR response time (in seconds) over Fluo-4 intensity in one representative CLL case. Addition of the stimulus is indicated by the arrow. (B and C) MFIs of phospho (p)-BTK (B) and p-ERK (C) in unstimulated and anti-IgM stimulated cells from 24 CLL cases. The histogram plot overlays below the graphs show p-BTK (B) and p-ERK (C) expression in the unstimulated and anti-IgM–stimulated cells from one representative CLL case; the light gray histograms correspond to unstained cells. (D) VLA-4 RO in primary CLL cells from 26 cases (all cases expressing >60% CD49d), pretreated or not with 1 µM ibrutinib and stimulated or not with 5 µg/ml anti-IgM. The reported VLA-4 RO values were calculated as described in Materials and methods and correspond to the presence of 10 nM LDV. (E) VLA-4 RO plotted versus LDV concentration using the sigmoidal dose–response equation with variable slope performed by GraphPad Prism software in cells untreated (left) or treated with ibrutinib (right) in one representative CLL case. The arrows indicate the shift from the control to the anti-IgM–stimulated condition at 10 nM LDV. EC 50 values for all conditions are indicated. R 2 corresponds to the coefficient of determination. (F) VLA-4 RO in primary CLL cells expressing CD49d between 10% and 29% ( n = 5) and between 30% and 60% ( n = 4), pretreated or not with 1 µM ibrutinib and stimulated or not with 5 µg/ml anti-IgM. (G) VLA-4 RO in T lymphocytes from 15 CLL cases in unstimulated and anti-IgM–stimulated conditions. The line graph on the right shows the VLA-4 RO plotted versus LDV concentration obtained in T-lymphocytes from one representative CLL case. (H) VLA-4 clusters in primary CLL cells from 3 cases treated as in (D) obtained on VCAM-1-coated slides. Quantitative clustering analysis was done in at least 50 individual cells for each condition by means of confocal microscopy (original magnification ×60). The three dot plots correspond to three different cases and show the cluster number for all analyzed cells in each condition. On the right, representative confocal microscopy images of VLA-4 clusters revealed by anti-CD49d mAbs, are shown. Bars, 2 µm. (I) Adhesion on VCAM-1 of primary CLL cells from 15 cases treated as in D. Cell adhesion was calculated as relative fold change obtained on VCAM-1 over BSA. Each experiment was run in triplicate. (J) Purified CLL cells from eight cases were stimulated or not with anti-IgM and perfused for 1 min at 0.5 dyn/cm 2 over immobilized VCAM-1. The data are expressed as the mean of frequencies of cells in direct contact with the substrate (tethering). Each experiment was run in triplicate. Data are presented as mean ± SEM. Individual symbols represent individual cases in all panels except H, which represents individual cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; n.s., not significant (Wilcoxon test in all panels except F, where a paired t test was applied).

Article Snippet: Slides were stained with primary anti-CD49d antibody (clone AHP1225; Bio-Rad) and Cy3-conjugated secondary anti–rabbit antibody.

Techniques: Activation Assay, In Vitro, Expressing, Concentration Assay, Software, Control, Confocal Microscopy, Purification

Journal: The Journal of Experimental Medicine

Article Title: Functional and clinical relevance of VLA-4 (CD49d/CD29) in ibrutinib-treated chronic lymphocytic leukemia

doi: 10.1084/jem.20171288

Figure Lengend Snippet: CD49d + and CD49d − CLL show different patterns of redistribution lymphocytosis during ibrutinib treatment. ALCs were collected pretreatment (day 0) and at different treatment time points (days 30, 60, 90, 120) in CLL cases from three different cohorts. (A) Kinetics of ALC (median values) in the IT cohort (left), NIH cohort (middle), and Mayo cohort (right); the gray and red symbols in each graph correspond to the pretreatment median ALC in CD49d − and CD49d + CLL, respectively. (B) Percent ALC change from baseline in CLL cases split according to CD49d expression (CD49d + , red; CD49d − , gray) from the three individual cohorts and the merging of all cohorts, shown as individual plots (above graphs) and as median values (below graphs). The number of patients included in each group are reported in parentheses; the black vertical lines indicate SEM. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001; n.s., not significant (Mann–Whitney test).

Article Snippet: Slides were stained with primary anti-CD49d antibody (clone AHP1225; Bio-Rad) and Cy3-conjugated secondary anti–rabbit antibody.

Techniques: Expressing, MANN-WHITNEY

Journal: The Journal of Experimental Medicine

Article Title: Functional and clinical relevance of VLA-4 (CD49d/CD29) in ibrutinib-treated chronic lymphocytic leukemia

doi: 10.1084/jem.20171288

Figure Lengend Snippet: VLA-4 expression decreases after ibrutinib therapy. (A) Flow-cytometric determination of CD49d, CD29, and IgM MFI in CLL cells collected at days 0 and 30 on ibrutinib. (B) Representative flow cytometry dot plot showing CXCR4 versus CD5 expression in gated CLL cells. The red gate identifies CXCR4 dim /CD5 bright population. (C) CD49d MFI in the CXCR4 dim /CD5 bright population of CLL cells collected at days 0 and 30 on ibrutinib. Data are presented as mean ± SEM. Individual symbols represent individual cases. *, P < 0.05; **, P < 0.01; n.s., not significant (Wilcoxon test).

Article Snippet: Slides were stained with primary anti-CD49d antibody (clone AHP1225; Bio-Rad) and Cy3-conjugated secondary anti–rabbit antibody.

Techniques: Expressing, Flow Cytometry

Journal: The Journal of Experimental Medicine

Article Title: Functional and clinical relevance of VLA-4 (CD49d/CD29) in ibrutinib-treated chronic lymphocytic leukemia

doi: 10.1084/jem.20171288

Figure Lengend Snippet: CD49d + and CD49d − CLL are characterized by different ibrutinib-induced nodal responses. LN dimension evaluation was performed pretreatment and at 12 mo of ibrutinib therapy, and the sum of the products of LN diameters (SPD) of up to five LN regions was calculated. (A) Waterfall plot showing the percent SPD change from baseline in CD49d + (red bars) or CD49d − (gray bars) CLL cases. (B–D) The box and whisker plot shows the median percent SPD change in CLL cases split according to CD49d expression (CD49d − , n = 23; gray box; CD49d + , n = 30; red box; B), TP53 disruption (no TP53 disruption, n = 24; blue box; TP53 disruption, n = 28; orange box; C), and IGHV mutations (UM, IGHV , n = 35; blue box; mutated [M] IGHV , n = 18; orange box; D). *, P < 0.05; n.s., not significant (Mann–Whitney test).

Article Snippet: Slides were stained with primary anti-CD49d antibody (clone AHP1225; Bio-Rad) and Cy3-conjugated secondary anti–rabbit antibody.

Techniques: Whisker Assay, Expressing, Disruption, MANN-WHITNEY

Journal: The Journal of Experimental Medicine

Article Title: Functional and clinical relevance of VLA-4 (CD49d/CD29) in ibrutinib-treated chronic lymphocytic leukemia

doi: 10.1084/jem.20171288

Figure Lengend Snippet: PFS of ibrutinib-treated CLL patients. (A-I) Kaplan–Meier estimates of PFS of CLL patients treated with ibrutinib and stratified by CD49d expression (A), TP53 disruption (B), IGHV mutational status (C), prior treatment (D), the association of CD49d expression and TP53 disruption (E), the association of CD49d expression and IGHV mutational status (F), CD49d expression in the context of RR CLL (G), TP53 disruption in the context of RR CLL (H), and IGHV mutational status in the context of RR CLL (I). The number of patients included in each group are reported in parentheses. *, P < 0.05; **, P < 0.01; n.s., not significant (log-rank test).

Article Snippet: Slides were stained with primary anti-CD49d antibody (clone AHP1225; Bio-Rad) and Cy3-conjugated secondary anti–rabbit antibody.

Techniques: Expressing, Disruption

Journal: The Journal of Experimental Medicine

Article Title: Functional and clinical relevance of VLA-4 (CD49d/CD29) in ibrutinib-treated chronic lymphocytic leukemia

doi: 10.1084/jem.20171288

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis of PFS in ibrutinib-treated CLL

Article Snippet: Slides were stained with primary anti-CD49d antibody (clone AHP1225; Bio-Rad) and Cy3-conjugated secondary anti–rabbit antibody.

Techniques:

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: A negative feedback loop between fibroadipogenic progenitors and muscle fibres involving endothelin promotes human muscle fibrosis

doi: 10.1002/jcsm.12974

Figure Lengend Snippet: Antibody panel used for CyTOF

Article Snippet: CD49d , 174Yb , 9F10 , Fluidigm , 3174018B.

Techniques: