Journal: Frontiers in Immunology

Article Title: Unveiling B7/CD28 family proteins in hepatocellular carcinoma: insights into communication and prognostic significance

doi: 10.3389/fimmu.2025.1583597

Figure Lengend Snippet: The expression pattern of B7H3 in tumor tissues. (A) The representative IHC images of Isotype control antibody and B7H3 antibody; (B) The representative IHC images of diffuse HCC and well-circumscribed HCC; (C) Representative multi-immunofluorescence images of AFP (green) and B7H3 (red). The nucleus is labeled with DAPI (blue). (D) Proportions of B7H3 on tumor cells (TCs), in stroma and in both. (E) The representative IHC images of B7H3 from serial sections. (F) OS and PFS for HCC patients based on the expression of B7H3 on tumor tissue. (G) OS and PFS for HCC patients based on the expression of B7H3 in the stroma and on tumor cells.

Article Snippet: In the end, the specimens were prepared by applying ProLongTM Diamond Antifade mountant with DAPI (G1236, Servicebio, Wuhan, China).

Techniques: Expressing, Control, Immunofluorescence, Labeling

Journal: The EMBO Journal

Article Title: Functional rescue of a disease-linked ERAD pathway mutation via alternative splicing

doi: 10.1038/s44318-026-00757-5

Figure Lengend Snippet: ( A ) The survival rate of WT, HET and KI pups at postnatal day 0 (p0). Observed vs. Expected * P = 0.013. ( B ) Body weight at p0. n = 7–9 mice/group. WT vs. KI, P = 0.94. ( C ) The photo of pups at postnatal 4 h (upper) and >12 h (lower). Scale bars: 1 cm. ( D ) Tissue morphology of SEL1L C141Y KI pups at P0, with tissue weight quantification shown below. n = 7–9 mice/group. Scale bars: 1 cm. WT vs. KI, Liver P = 0.45, Brain P = 0.65, Thymus P = 0.14, Spleen P = 0.16. ( E ) Morphology of postprandial stomach in the P0 WT and KI pup, with quantification shown on the right. n = 7–9 mice/group. Scale bars : 1 cm. WT vs. KI, **** P = 0.000013. ( F ) The survival curve of WT, HET and KI pups after birth. n = 17–41 mice/group. WT vs. HET, P = 0.86; WT vs. KI, **** P < 0.0001. ( G ) Body weight growth of male and female WT, HET, KI’ mice for postnatal week 1 to week 10. n = 3–20 mice/group. ( H ) Quantification of B cells as a percentage of CD45⁺ peripheral blood mononuclear cells (PBMCs) at p0 by flow cytometry in Fig. . n = 5–10 mice/group. WT vs. KI, *** P = 0.0002; WT vs. KI’, P = 0.63; KI vs. KI’, ** P = 0.0091. ( I , J ) Hematoxylin and eosin (H&E) staining and Immunofluorescence of CD19 + B cells in p0 mouse spleens from SEL1L C141Y KI pups at p0, with quantification shown in ( J ). Red, CD19; Blue, DAPI. n = 3 mice/group. Scale bars: 50 μm for zoom-out photo, 20 μm for zoom-in photo and immunofluorescence figure. *** P = 0.0026. Data are represented as means ± SEM. n.s. not significant. * P < 0.05; ** P < 0.01; *** P < 0.001, **** P < 0.0001, by Chi-square test for ( A ); two-tailed t test for ( B , D , E , J ); Mantel–Cox test for ( F ); two-way ANOVA followed by Tukey’s multiple-comparisons test for ( G ); one-way ANOVA followed by Tukey’s post hoc test for ( H ). .

Article Snippet: ProLongTM Diamond Antifade Mountant with DAPI , Fisher Scientific , P36971.

Techniques: Flow Cytometry, Staining, Immunofluorescence, Two Tailed Test

Journal: The EMBO Journal

Article Title: Functional rescue of a disease-linked ERAD pathway mutation via alternative splicing

doi: 10.1038/s44318-026-00757-5

Figure Lengend Snippet: ( A – C ) Western blot analysis of SEL1L protein expression ( A ), HRD1 and ERAD substrate IRE1α ( B ) in mouse brains or cortex at indicated ages, with quantification in ( C ). n = 3–4 mice/group. SEL1L: WT vs. HET, P = 0.057; WT vs. KI, *** P = 0.0006; HRD1: WT vs. HET, P = 0.52; WT vs. KI, ** P = 0.0017; IRE1α: WT vs. HET, P = 0.99; WT vs. KI, **** P < 0.0001. ( D , E ) Western blot analysis ( D ) of ERAD protein and ERAD substrates from WT and KI’ mice brain with quantification shown in ( E ). n = 3–4 mice/group. SEL1L: WT vs. KI’, *** P = 0.0009; HRD1: WT vs. KI’, P = 0.34; IRE1α: WT vs. KI’, P = 0.28. ( F ) Phos-tag gel analysis of IRE1α phosphorylation in p0 WT, HET and KI brains. n = 3–4 mice/group. ( G ) RT-PCR analysis of Xbp1 mRNA splicing of p0 WT, KI, and KI’ brains with quantification in Fig. . n = 3–6 mice/group, 3–4 for positive controls. u, unspliced. s, spliced. ( H , I ) Immunofluorescence ( H ) of SATB2+ Layer II/III cells and the CTIP2+ Layer V in the cortex of p0 WT, KI and KI’ mice, with zoom-in region showing SATB2+ cells below Layer V and with quantification shown in ( I ). n = 3–6 mice/group. Green, CTIP2; Red, SATB2; Blue, DAPI. WT vs. KI, * P = 0.046; WT vs. KI’, * P = 0.032; KI vs. KI’, *** P = 0.0007. Data are represented as means ± SEM. n.s. not significant. * P < 0.05; ** P < 0.01; *** P < 0.001, **** P < 0.0001, by one-way ANOVA followed by Tukey’s post hoc test for ( C , I ); two-tailed t test for ( E ). .

Article Snippet: ProLongTM Diamond Antifade Mountant with DAPI , Fisher Scientific , P36971.

Techniques: Western Blot, Expressing, Phospho-proteomics, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Two Tailed Test

Journal: Parasites & Vectors

Article Title: A polo-like kinase modulates cytokinesis and flagella biogenesis in Giardia lamblia

doi: 10.1186/s13071-021-04687-5

Figure Lengend Snippet: Expression and localization of G. lamblia PLK1 (GlPLK) in G. lamblia -expressing hemagglutinin (HA)-tagged GlPLK. a A schematic diagram of plasmid pGlPLK.neo. HA-tagged GlPLK was expressed from its own promoter, P glplk . Transfected trophozoites were selected by neomycin resistance conferred by the neo gene expressed by the P glggi promoter, a strong promoter of the γ-giardin gene. As a control, Giardia trophozoites were also transfected with pKS-3HA.neo, a vector control. b Western blotting to examine the expression of HA-tagged GlPLK. Extracts were prepared from G. lamblia containing empty vector (lane 1) or pGlPLK.neo (lane 2) and incubated with monoclonal mouse anti-HA antibodies. Membranes were first incubated in stripping buffer and then reacted with polyclonal rat antibodies specific to protein disulfide isomerase 1 (PDI1) of G. lamblia . c Localization of GlPLK. Giardia lamblia expressing HA-tagged GlPLK was probed with mouse anti-HA antibodies. The cells were then incubated with Alexa Fluor 488-conjugated anti-mouse IgG. Slides were mounted with ProLong™ Gold Antifade Mountant with the fluorescent stain DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope. Scale bars: 2 µm. d Co-localization of GlPLK and α-tubulin in G. lamblia . Giardia cells expressing HA-tagged GlPLK were probed with rat anti-HA antibodies and mouse anti-acetylated-α-tubulin monoclonal antibodies. e Co-localization of GlPLK and G. lamblia centrin (GlCentrin) in G. lamblia . Cells were reacted with mouse anti-HA antibodies and rat anti-GlCentrin polyclonal antibodies. Cells were then incubated with Alexa Fluor 555-conjugated anti-rat IgG and Alexa Fluor 488-conjugated anti-mouse IgG. A differential interference contrast image was acquired to show cell morphology. Scale bars: 2 μm. DIC Differential interference contrast

Article Snippet: Slides were mounted with ProLongTM Gold Antifade Mountant with DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope.

Techniques: Expressing, Plasmid Preparation, Transfection, Control, Western Blot, Incubation, Stripping Membranes, Staining, Laser-Scanning Microscopy, Bioprocessing

Journal: Parasites & Vectors

Article Title: A polo-like kinase modulates cytokinesis and flagella biogenesis in Giardia lamblia

doi: 10.1186/s13071-021-04687-5

Figure Lengend Snippet: Expression and localization of truncated GlPLKs in G. lamblia . a A schematic diagram of plasmids pGlPLKKDL.neo and pGlPLKPBD.neo. Two truncated GlPLK proteins are expressed in an HA-tagged form from their own promoter, P glplk . Plasmid pGlPLKKDL encodes GlPLK with the KD and linker region, whereas pGlPLKPBD contains DNA coding for the polo-box domains (PBDs) of GlPLK. Plasmid pKS-3HA.neo was transfected into Giardia trophozoites as a control. b Western blotting to examine the expression of HA-tagged truncated GlPLKs. Extracts were prepared from G. lamblia containing empty vector (lane 1), pGlPLKKDL.neo (lane 2), or pGlPLKPBD.neo (lane 3), and incubated with monoclonal mouse anti-HA antibodies. Membranes were incubated in stripping buffer, and then reacted with polyclonal rat antibodies specific to GlPDI1. c Co-localization of GlPLK-KDL with α-tubulin ( i ) or GlCentrin ( ii , iii ). Giardia lamblia cells expressing HA-tagged truncated GlPLK-KDL were probed with rat anti-HA antibodies and mouse anti-acetylated-α-tubulin monoclonal antibodies. Otherwise, these cells were reacted with rat anti-GlCentrin polyclonal antibodies instead of anti-acetylated-α-tubulin monoclonal antibodies. Panel iii is an extended view of panel ii . Incorrectly positioned basal bodies are indicated with white arrows. d Co-localization of GlPLK-PBD with α-tubulin ( i ) or GlCentrin ( ii , iii ). G. lamblia cells expressing HA-tagged truncated GlPLK-PBD were probed with rat or mouse anti-HA antibodies along either with mouse anti-acetylated-α-tubulin antibodies ( i ), or rat anti-GlCentrin polyclonal antibodies ( ii , iii ), respectively. Panel iii is an extended view of panel ii . The cells were then incubated with Alexa Fluor 488-conjugated anti-rat IgG and Alexa Fluor 568-conjugated anti-mouse IgG (α-tubulin co-localization) or Alexa Fluor 555-conjugated anti-rat IgG and Alexa Fluor 488-conjugated anti-mouse IgG (for centrin co-localization). Slides were mounted with ProLong™ Gold Antifade Mountant with DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope. A differential interference contrast image was acquired to show cell morphology. Scale bars: 2 μm

Article Snippet: Slides were mounted with ProLongTM Gold Antifade Mountant with DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope.

Techniques: Expressing, Plasmid Preparation, Transfection, Control, Western Blot, Incubation, Stripping Membranes, Bioprocessing, Laser-Scanning Microscopy

Journal: Parasites & Vectors

Article Title: A polo-like kinase modulates cytokinesis and flagella biogenesis in Giardia lamblia

doi: 10.1186/s13071-021-04687-5

Figure Lengend Snippet: Expression and localization of phosphorylated GlPLK in G. lamblia . a Giardia lamblia cells expressing HA-tagged GlPLK in the interphase (lane 1), G1/S phase (lane 2), and G2/M phase (lane 3). Western blot analysis. Extracts of these cells were probed with anti-phospho-PLK, anti-HA, anti-GlPLK or anti-GlPDI1 antibodies. Levels of phospho-GlPLK-HA, phospho-GlPLK, GlPLK-HA and GlPLK were normalized to GlPDI1, a protein loading control. Amounts of these GlPLK proteins are expressed as relative values to those observed in the interphase cells. The presented western blot is a representative of three independent experiments, and averages of three experiments are presented as a bar graph. b Localization of GlPLK and phosphorylated GlPLK in Giardia . G. lamblia expressing HA-tagged GlPLK were incubated with antibodies specific to the phosphorylated form of PLK (1:100) along with anti-HA antibodies. These cells were then reacted with anti-Alexa Fluor 568-conjugated anti-mouse IgG and Alexa Fluor 488-conjugated anti-rat IgG. Slides were mounted with ProLong™ Gold Antifade Mountant with DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope. Scale bars: 2 µm

Article Snippet: Slides were mounted with ProLongTM Gold Antifade Mountant with DAPI, and then examined with a Zeiss LSM700 inverted confocal laser scanning microscope.

Techniques: Expressing, Western Blot, Control, Incubation, Laser-Scanning Microscopy