Journal: Cells

Article Title: A Mouse Model to Study the Pathogenesis of γ-herpesviral Infections in Germinal Center B Cells.

doi: 10.3390/cells12242780

Figure Lengend Snippet: Figure 5. Virally induced constitutively active CD30 signaling results in more Cre-reporter+ cells within B cells. CAR//Cγ1-cre mice (dots) and LC30//Cγ1-cre mice (squares) were i.p. infected with MHV-68 hNGFR and splenic cells were analyzed 14 days post-infection. (A) Breeding scheme of LC30//Cγ1cre mice. The excision of the loxP-flanked stop cassette was followed by hCD2 reporter expression (Cre-reporter) and ligand-independent constitutively active CD30 signaling. (B) The splenic

Article Snippet: The CD19-PeVio770 (REA749), CD38-PeVio770 and human CD2 (hCD2)-PeVio770 (LT2) antibodies were all purchased from Miltenyi Biotech (Bergisch Gladbach, Germany).

Techniques: Infection, Expressing

Journal: bioRxiv

Article Title: Ångström Resolution with Flow Immunofluorescence Localization Microscopy (FILM)

doi: 10.64898/2025.12.16.694546

Figure Lengend Snippet: (A–C) FILM acquisition of binder probes that recognize CD2, ICAM-1 (CD54) in the pSMAC, and CD45 in the dSMAC zones (10 nm pixels with 3-pixel Gaussian blurring). For CD2 and CD45, APC-conjugated antibodies were used; for ICAM-1, a nanobody Fab conjugated with Vio ® 667 was used. Cyan boxed region shows the IF image (103.17 nm pixels) corresponding to the white boxed region. (D–F) Zoom-in of the respective boxed regions for the panels to the left. Localizations are shown as peak-normalized 2D Gaussians with color-coded precision according to the colorbar. (G–I) Zoom-in of the respective boxed regions for the panels to the immediate left. Localizations are shown as crosses (indicating ±1-σ uncertainty in x and y ) with color-coded precision according to the colorbar.

Article Snippet: α-tubulin (REA1136 REAfinityTM, h, APC, #130-119-541, Miltenyi Biotec), CD2 (REA972 REAfinityTM, h, APC, #130-116-150, Miltenyi Biotec), CD20 (REA780 REAfinityTM, h, APC, #130-111-339, Miltenyi Biotec), CD28 (REA612 REAfinityTM, h, APC, #130-118-343, Miltenyi Biotec), CD45 (REA1023 REAfinityTM, h/nhp, APC, #130-117-191, Miltenyi Biotec), CD54/ICAM-1 (REA266 REAfinityTM, h, APC, #130-120-711, Miltenyi Biotec), CD81 (REA513 REAfinityTM, h, APC, #130-119-787, Miltenyi Biotec), CD107a/LAMP1 (REA792 REAfinityTM, h, APC, #130-111-847, Miltenyi Biotec), IgD (REA740 REAfinityTM, h, APC, #130-110-644, Miltenyi Biotec), IgM (REAL689 REAdye_leaseTM, h, PE, #130-125-786, Miltenyi Biotec), Pan-Cytokeratin (REA1141 REAfinityTM, h, APC, #130-120-096, Miltenyi Biotec), β-catenin (REA480 REAfinityTM, h, APC, #130-124-444, Miltenyi Biotec), TCRα/β (REA652 REAfinityTM, h, APC, #130-113-535, Miltenyi Biotec), TOM22 (REA1185 REAfinityTM, APC, h/ms, #130-122-079, Miltenyi Biotec), DAPI Staining Solution (#130-111-570, Miltenyi Biotec), and ATTO 643 phalloidin (ATT-AD643-81, ATTO-TEC).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Exploration of the mechanism of Taohong Siwu Decoction for the treatment of ischemic stroke based on CCL2/CCR2 axis

doi: 10.3389/fphar.2024.1428572

Figure Lengend Snippet: Expression of CD86 (M1) and CD206 (M2). (A) : Expression of CD86 (M1); (B) : Expression of CD206 (M2). # P < 0.05, ## P < 0.01 vs. sham; * p < 0.05, ** p < 0.01 vs. MCAO/R.

Article Snippet: A11042), were both purchased from MultiSciences, APC anti-Human CD206 (LW1218) was purchased from Elabscience, human TNF-α (July 2023), Human matrix metalloproteinase 9 (MMP-9) (July 2023), human IL-6 (July 2023), human IL-4 (July 2023), rat TNF-α (May 2023), rat MMP-9 (May 2023), rat IL-6 (May 2023), rat IL-4 (May 2023) were purchased from Quanzhou Ruixin Biotechnology Co., Ltd., claudin-5 (cat.no.

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: Identification of Novel Prognostic Biomarkers Relevant to Immune Infiltration in Lung Adenocarcinoma

doi: 10.3389/fgene.2022.863796

Figure Lengend Snippet: The correlations between PSTPIP1/PILRA and immune biomarkers in LUAD. Scatter plots showing the correlation between (A) PSTPIP1 and PD-L1, (B) PSTPIP1 and IFN-γ, (C) PILRA and PD-L1, and (D) PILRA and IFN-γ in the LUAD samples ( n = 515). Violin plots showing the expression of (E) PSTPIP1 and (F) PILRA in the LUAD samples with different degrees of differentiation (including 167 with poor differentiation, 209 with moderate differentiation, and 60 with well differentiation). ns indicates p ≥ 0.05, * p < 0.05.

Article Snippet: Next, the slices were incubated with the primary antibodies anti-PSTPIP1 (11951-1-AP, rabbit, polyclonal, dilution 1:50, Proteintech, Wuhan, China), anti-PILRA (orb38981, rabbit, polyclonal, dilution 1:200, Biorbyt, Cambridge, United Kingdom), and PBS (blank control) overnight at 4°C, washed three times with PBS, and incubated with a horseradish peroxidase (HRP)–conjugated secondary antibody (ab205718, Abcam, Cambridge, United Kingdom) for 30 min.

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: Identification of Novel Prognostic Biomarkers Relevant to Immune Infiltration in Lung Adenocarcinoma

doi: 10.3389/fgene.2022.863796

Figure Lengend Snippet: The influence of PSTPIP1 and PILRA on immune infiltration and prognosis in LUAD. Violin plots showing (A) the immune score in samples with low or high expression of PSTPIP1 and PILRA. GSVA-derived clustering heat maps of differentially infiltrated immune cell populations between the high and low expression groups of (B) PSTPIP1 and (C) PILRA. Only lymphocytes with log(fold change) > 0.2 are shown. The influence of (D) PSTPIP1 and (E) PILRA on the overall survival time of LUAD patients. The yellow line indicates samples with highly expressed genes and the blue line indicates samples with lowly expressed genes. Violin plots showing (F) the expression of PSTPIP1 and PILRA in the LUAD patients with different responses to nivolumab (including five responders and five nonresponders) and (G) the expression of PSTPIP1 in SKCM mice with different responses to anti–PD-L1 treatment (including 27 responders and 23 nonresponders). ns indicates p ≥ 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Next, the slices were incubated with the primary antibodies anti-PSTPIP1 (11951-1-AP, rabbit, polyclonal, dilution 1:50, Proteintech, Wuhan, China), anti-PILRA (orb38981, rabbit, polyclonal, dilution 1:200, Biorbyt, Cambridge, United Kingdom), and PBS (blank control) overnight at 4°C, washed three times with PBS, and incubated with a horseradish peroxidase (HRP)–conjugated secondary antibody (ab205718, Abcam, Cambridge, United Kingdom) for 30 min.

Techniques: Expressing, Derivative Assay

Journal: Frontiers in Genetics

Article Title: Identification of Novel Prognostic Biomarkers Relevant to Immune Infiltration in Lung Adenocarcinoma

doi: 10.3389/fgene.2022.863796

Figure Lengend Snippet: Histopathological examples of PSTPIP1/PILRA expression and tumor-infiltrating lymphocytes (TILs) in LUAD. (A) PSTPIP1 and PILRA expression is demonstrated by brown staining, and the bar plot shows the average optical density of PSTPIP1 and PILRA in the PD-L1–positive or PD-L1–negative LUAD tissue samples. (B) The TILs are displayed as purple spots in hematoxylin and eosin staining, and bar plots show the stromal TILs in tissue samples with high and low PSTPIP1/PILRA expressions. * p < 0.05.

Article Snippet: Next, the slices were incubated with the primary antibodies anti-PSTPIP1 (11951-1-AP, rabbit, polyclonal, dilution 1:50, Proteintech, Wuhan, China), anti-PILRA (orb38981, rabbit, polyclonal, dilution 1:200, Biorbyt, Cambridge, United Kingdom), and PBS (blank control) overnight at 4°C, washed three times with PBS, and incubated with a horseradish peroxidase (HRP)–conjugated secondary antibody (ab205718, Abcam, Cambridge, United Kingdom) for 30 min.

Techniques: Expressing, Staining

Journal: Nature Communications

Article Title: Noncanonical function of folate through folate receptor 1 during neural tube formation

doi: 10.1038/s41467-024-45775-1

Figure Lengend Snippet: a Neural plate stage Xenopus laevis embryos were processed for co-immunoprecipitation (IP) assays. Example of Western blot assay from immunoprecipitates with FOLR1 or GFP (control) antibodies and probed for CD2AP or FOLR1. Similar results were observed in N = 3 independent experiments. b Neural plate stage Xenopus laevis embryos were fixed and processed for immunostaining. Images are transverse single z-sections of immunostained neural plate showing apical colocalization of phospho-CD2AP (p-CD2AP) and p-c-Cbl with C-cadherin. Scale bar, 10 μm. Similar results were observed in N = 3 independent experiments. c Two-cell stage Xenopus laevis embryos were bilaterally microinjected with 9.9 pmol of Control-morpholino (Control, Control-MO), 2.6 pmol CD2AP-MO1 (CD2AP KD1) or 7.4–9.9 pmol CD2AP-MO2 (CD2AP KD2/KD) per blastomere until neural tube closed in control embryos, when they were fixed and photomicrographed. Examples of whole embryos in each group. Arrowheads indicate open neural tube (neural tube defect, NTD). Numbers are embryos presenting closed (green) or open (purple, NTD) neural tube. Bar graph represents proportion of phenotypes in each group. d Two-cell stage Xenopus laevis embryos were unilaterally microinjected with 9.9 pmol Control-MO (Control) and 7.4–9.9 pmol CD2AP-MO2 (CD2AP KD) along with GFP and mCherry mRNA, respectively, and allowed to develop until they reached mid-neural plate stages, when they were fixed and processed for immunostaining. Image is a transverse section of the neural plate, immunostained for GFP (Control), mCherry (CD2AP KD) and C-cadherin. Double arrows indicate apical surface length of medial superficial Control (white) and CD2AP KD (magenta) neural plate cells. Scale bar, 20 μm. Graph shows individual and mean ± SD apical length of superficial neural plate cells per embryo, n of cells = 75 in each half of the neural plate, N of embryos = 4. **** p < 0.0001, two-tailed paired t -test. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used were: anti-phospho-tyr-4,8,10 CD2AP, 1:500 (Rockland, cat. # 600-401-J96), anti-phospho-tyr-674 c-Cbl, 1:500 (MyBioSource, cat. # MBS820886), anti-C-cadherin, 1:100 (Developmental Studies Hybridoma Bank, cat. # 6B6), anti-EEA1, 1:1000 (Origene, cat. # AB0006-200), anti-ubiquitin, 1:500 (Stress Marq, cat. # SPC-119B), anti-Rab7, 1:500 (Cell Signaling, cat. # 9367), anti-LAMP1, 1:500 (Abcam, cat. # ab24170), anti-GFP, 1:750 (Abcam, cat. # ab13970), anti-mCherry, 1:750 (Biorbyt, cat. # orb11618), anti-SOX2, 1:300 (Cat # AF2018, R&D Systems), anti-α-tubulin, 1:500 (Abcam, cat. # ab15246).

Techniques: Immunoprecipitation, Western Blot, Control, Immunostaining, Two Tailed Test

Journal: Nature Communications

Article Title: Noncanonical function of folate through folate receptor 1 during neural tube formation

doi: 10.1038/s41467-024-45775-1

Figure Lengend Snippet: a Two-cell stage Xenopus laevis embryos were bilaterally microinjected with hEEA1-GFP and membrane mCherry mRNAs and unilaterally microinjected with 9.9 pmol CD2AP-MO (CD2AP KD) per blastomere along with fluorescent tracer and allowed to develop until they reached early neural plate stages (stage 13-14), when they were time-lapse imaged with an acquisition rate of 1 frame/6 min. Image is maximum intensity projection of single time frame. Dashed line indicates border between morpholino-injected and wild-type (WT) neural plate. Inset shows neural plate injected side showing tracer in blue. Graphs show distribution between both halves of the neural plate (in %) of the number of EEA1-GFP vesicles and area fraction of labeled endosomes per neural plate cell surface. Two-tailed paired t -test, n = 28 cells analyzed in each group from N = 5 embryos per group. Scale bar, 20 μm. b Two-cell stage Xenopus laevis embryos were bilaterally microinjected with 7.4 pmol Control-MO (Control) or CD2AP-MO (CD2AP KD) per blastomere and allowed to grow until they reached mid-neural plate stages (stage 15–17) when neural plate was dissected and processed for Western blot assays. Image is an example of Western blot assay. Graph shows individual and mean ± SD percent of optical density (OD) for C-cadherin immunoblot band normalized with GAPDH protein band OD and compared to controls. Two-tailed ratio t -test, n = 28 and 24 neural plates for Control and CD2AP KD groups, respectively, N = 5 independent experiments. In ( a , b ), * p < 0.05, ** p < 0.01, *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used were: anti-phospho-tyr-4,8,10 CD2AP, 1:500 (Rockland, cat. # 600-401-J96), anti-phospho-tyr-674 c-Cbl, 1:500 (MyBioSource, cat. # MBS820886), anti-C-cadherin, 1:100 (Developmental Studies Hybridoma Bank, cat. # 6B6), anti-EEA1, 1:1000 (Origene, cat. # AB0006-200), anti-ubiquitin, 1:500 (Stress Marq, cat. # SPC-119B), anti-Rab7, 1:500 (Cell Signaling, cat. # 9367), anti-LAMP1, 1:500 (Abcam, cat. # ab24170), anti-GFP, 1:750 (Abcam, cat. # ab13970), anti-mCherry, 1:750 (Biorbyt, cat. # orb11618), anti-SOX2, 1:300 (Cat # AF2018, R&D Systems), anti-α-tubulin, 1:500 (Abcam, cat. # ab15246).

Techniques: Membrane, Injection, Labeling, Two Tailed Test, Control, Western Blot

Journal: Nature Communications

Article Title: Noncanonical function of folate through folate receptor 1 during neural tube formation

doi: 10.1038/s41467-024-45775-1

Figure Lengend Snippet: Two-cell stage Xenopus laevis embryos were microinjected with 14.8 pmol Control-morpholino (MO, Control, a , b ), 3.2 pmol FOLR1-MO (FOLR1 KD, a ) or 14.8 pmol CD2AP-MO2 (CD2AP KD, b ) per embryo and incubated with saline or proteasome and lysosome inhibitors at the end of gastrulation (stage 12) until neural plate stages (stage 17) when they were processed for Western blot assays. Images are examples of Western blot assays. Graphs show individual and mean ± SD percent of optical density (OD) for CD2AP ( a ) or FOLR1 ( b ) immunoblot band normalized with GAPDH protein band OD and compared to controls. In ( a ), n = 34 and 42 neural plates for Control and FOLR1 KD, respectively, N = 7 independent experiments; n = 20 and 26 neural plates for Control+inhibitors and FOLR1 KD+inhibitors groups, respectively, N = 3 independent experiments. In ( b ) n = 16 and 20 for Control and CD2AP KD groups, respectively and n = 16 and 18 neural plates for Control+inhibitors and CD2AP KD+inhibitors, respectively, N = 3 independent experiments. In ( a , b ) * p < 0.05, *** p < 0.001, ns: not significant, two-tailed ratio t -test. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used were: anti-phospho-tyr-4,8,10 CD2AP, 1:500 (Rockland, cat. # 600-401-J96), anti-phospho-tyr-674 c-Cbl, 1:500 (MyBioSource, cat. # MBS820886), anti-C-cadherin, 1:100 (Developmental Studies Hybridoma Bank, cat. # 6B6), anti-EEA1, 1:1000 (Origene, cat. # AB0006-200), anti-ubiquitin, 1:500 (Stress Marq, cat. # SPC-119B), anti-Rab7, 1:500 (Cell Signaling, cat. # 9367), anti-LAMP1, 1:500 (Abcam, cat. # ab24170), anti-GFP, 1:750 (Abcam, cat. # ab13970), anti-mCherry, 1:750 (Biorbyt, cat. # orb11618), anti-SOX2, 1:300 (Cat # AF2018, R&D Systems), anti-α-tubulin, 1:500 (Abcam, cat. # ab15246).

Techniques: Control, Incubation, Saline, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Noncanonical function of folate through folate receptor 1 during neural tube formation

doi: 10.1038/s41467-024-45775-1

Figure Lengend Snippet: a – c Neural plate from mid neural plate stage Xenopus laevis embryos was dissected and dissociated cells plated in vitro. After 2 h, cells were loaded with the Ca 2+ sensor Fluo4-AM and time-lapse imaged. a Example of 1-h recording of neural plate cell Ca 2+ activity. b , c Folinic acid ( b , c ), folic acid ( c ) or vehicle was added to neural plate cells in culture during time-lapse imaging and the Ca 2+ response was recorded in the first minute post addition. b Example of acute transient elicited by 100 μM folinic acid. c Graph shows mean ± SEM folinic- or folic acid-responsive neural plate cells compared to total number of cells with spontaneous Ca 2+ transients in 30 min recording, N = 3 independent experiments. Two-tailed one sample t and Wilcoxon test. d Two-cell stage embryos were bilaterally microinjected with GCaMP6s mRNA and grown until early and mid-neural plate stages when they were time-lapse imaged before and after addition of vehicle or 300 μM folinic acid. Image shows example of embryo with cells exhibiting Ca 2+ transients indicated with circles. Graph shows individual and mean ± SD percent change in Ca 2+ transient frequency before and after addition of vehicle or folinic acid, n = 4, 5, 6 and 7 embryos for Early-Vehicle, Early-Folinic acid, Mid-Vehicle and Mid-Folinic acid groups, respectively. One-sample two-tailed t -test, compared to hypothetical value of 100%. e – h Two-cell stage embryos were bilaterally microinjected with GCaMP6s mRNA ( e – h ) and unilaterally microinjected with 9.9 pmol FOLR1-MO1 (FOLR1 KD1/KD) or 1.6 pmol FOLR1-MO2 (FOLR1 KD2) per blastomere ( e ) and grown until mid-neural plate stages when they were time-lapse imaged. Image in ( e ) shows example of embryo with cells exhibiting Ca 2+ transients indicated with circles in WT and FOLR1 KD1 halves. Graphs show individual Ca 2+ transient frequency (transients/5 min) in WT and FOLR1 KD1 or KD2 halves ( e , n = 6 embryos) and in WT embryos before and after addition of vehicle ( f , n = 7 embryos), 50 μM folic acid ( g , n = 6 embryos), Na + and voltage-gated Ca 2+ channel blockers (VGC block : 0.02% tricaine+10 μM nitrendipine+25 μM TTA-2, h , n = 5 embryos), or a mix of folic acid and VGC block ( h , n = 5 embryos). Two-tailed paired t -test ( e – g ) and 1-way ANOVA-Tukey multiple comparisons test ( h ). In ( c – h ), * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant. i Model of FOLR1 and CD2AP regulation of neural tube formation. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies used were: anti-phospho-tyr-4,8,10 CD2AP, 1:500 (Rockland, cat. # 600-401-J96), anti-phospho-tyr-674 c-Cbl, 1:500 (MyBioSource, cat. # MBS820886), anti-C-cadherin, 1:100 (Developmental Studies Hybridoma Bank, cat. # 6B6), anti-EEA1, 1:1000 (Origene, cat. # AB0006-200), anti-ubiquitin, 1:500 (Stress Marq, cat. # SPC-119B), anti-Rab7, 1:500 (Cell Signaling, cat. # 9367), anti-LAMP1, 1:500 (Abcam, cat. # ab24170), anti-GFP, 1:750 (Abcam, cat. # ab13970), anti-mCherry, 1:750 (Biorbyt, cat. # orb11618), anti-SOX2, 1:300 (Cat # AF2018, R&D Systems), anti-α-tubulin, 1:500 (Abcam, cat. # ab15246).

Techniques: In Vitro, Activity Assay, Imaging, Two Tailed Test, Blocking Assay