Journal:

Article Title: Temporally Regulated Neural Crest Transcription Factors Distinguish Neuroectodermal Tumors of Varying Malignancy and Differentiation 1

doi:

Figure Lengend Snippet: Figure 2

Article Snippet: Mouse monoclonal antibodies to AP-2α (3B5; developed by Dr. Trevor Williams) and PAX7 (PAX7; developed by Dr. Atsushi Kawakami) were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by The University of Iowa (Iowa City, IA).

Techniques:

Journal:

Article Title: Temporally Regulated Neural Crest Transcription Factors Distinguish Neuroectodermal Tumors of Varying Malignancy and Differentiation 1

doi:

Figure Lengend Snippet: Graph 2

Article Snippet: Mouse monoclonal antibodies to AP-2α (3B5; developed by Dr. Trevor Williams) and PAX7 (PAX7; developed by Dr. Atsushi Kawakami) were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by The University of Iowa (Iowa City, IA).

Techniques:

Journal:

Article Title: TBX5 is required for embryonic cardiac cell cycle progression

doi: 10.1242/dev.02420

Figure Lengend Snippet: (A) RT-PCR analysis of the expression of heart-specific isoforms of MHC, troponin and tropomyosin; and skeletal muscle-specific genes MyoD and muscle actin, throughout early and mid-tadpole stages of development in CMO (‘C’) and T5MO (‘T’) stage-matched embryos. All samples are derived from a single batch of eggs, and identical results were achieved in at least two independent sets of experiments for each marker. EF1-Alpha was used as a loading control for all RT-PCR reactions. (B–I) Images depicting embryos injected with (B–E) CMO, or (F–I) T5MO and immunostained for Tmy, showing delayed expression of Tmy in the hearts of T5MO embryos. Shown are representative sibling embryos imaged at the indicated stages. White arrows denote expression of Tmy within the heart. (J–Q) Images of living cardiac actin:GFP transgenic embryos, showing a delay in the onset of cardiac actin expression in the heart. Representative sibling embryos obtained from a single batch of embryos were injected with (J–M) CMO or (N–Q) T5MO and imaged at the indicated stages. Shown is a representative pair of embryos, while identical results were observed in over 50 embryos. White arrows denote expression of GFP within the heart field.

Article Snippet: The antibodies against tropomyosin and cardiac troponin (developed by J.-C. Lin), and fibrillin (developed by C. D. Little) were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Derivative Assay, Marker, Control, Injection, Transgenic Assay

Journal:

Article Title: TBX5 is required for embryonic cardiac cell cycle progression

doi: 10.1242/dev.02420

Figure Lengend Snippet: Cardiomyocyte structure in transverse sections through the hearts of (A,C,E,G) CMO or (B,D,F,H) T5MO stage 37 embryos, as detected by immunostaining for (A,B) cardiac troponin T (cTNT), (C,D) MHC, (E,F) actin or (G,H) Tmy. (I,K,M) Stage 37 CMO or (J,L,N) T5MO embryos double-immunostained for tropomyosin (green) and (I,J) fibronectin, (K,L) fibrillin or (M,N) β-catenin, all shown in red. Note increase in fibrillin staining on the walls of the chamber of T5MO hearts relative to CMO (compare panel K to L, white arrows) and ectoptic expression of fibronectin, shown by white arrow, in the dorsal portion of the heart in panel J relative to panel I. (O,P) High magnification confocal images of hearts from (O) CMO or (P) T5MO stage 37 embryos. Note that formation of organized cardiac muscle bundles in T5MO hearts is limited to a single cluster adjacent to the cardiac lumen. (Q–S) Representative transmission electron micrographs of transverse images of stage 37 embryos taken from (Q) CMO cardiac tissue or (R) T5MO cardiac tissue adjacent to the pericardial cavity and (S) T5MO cardiac tissue adjacent to the cardiac lumen. Cardiac muscle fibrils are shown pseudo-colored in yellow. Note that sarcomeres in T5MO hearts can only be identified adjacent to the cardiac lumen (compare R with S) and only found in concentric arrays. By contrast, CMO-derived hearts show both longitudinal and concentric arrays (compare Q with S). High-magnification TEM images reveal the ultrastructures of (T) CMO and (U) T5MO cardiac sarcomeres. Arrows denote A-bands. Note the smaller, non-continuous A-bands in the T5MO-derived sarcomeres (U). (V) Traces of the heart sections from CMO and T5MO embryos imaged by TEM are depicted schematically. Yellow circles represent the location of TEM imaging. Scale bars: 50 μm in A–N; 2 μm in Q–S; 0.2 μm in T,U.

Article Snippet: The antibodies against tropomyosin and cardiac troponin (developed by J.-C. Lin), and fibrillin (developed by C. D. Little) were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.

Techniques: Immunostaining, Staining, Expressing, Transmission Assay, Derivative Assay, Imaging

Journal:

Article Title: TBX5 is required for embryonic cardiac cell cycle progression

doi: 10.1242/dev.02420

Figure Lengend Snippet: The overall morphology of stage 40 (A) uninjected embryos or (B,C) embryos injected with increasing amounts of Tbx5 RNA, as indicated. Arrows denote the location of the heart. (D–G) Whole-mount in-situ hybridization showing expression of (D,E) Nkx2.5 and (F,G) myosin light chain (MLC) in (D,F) uninjected stage 37 embryos and (E,G) stage-matched embryos injected with 1 ng of Tbx5 RNA. (H) Tbx5 misexpression leads to an increase in the cardiac mitotic index at stage 37. Mitotic index was calculated as the percentage of cardiac cells labeled with pH3. The data represents the mean of at least three different embryos. Error bars denote the standard deviation, and * denotes a statistically significant difference between Tbx5-injected and control embryos (at P<0.05). (I) Mitotic index for sections of the neural tube corresponding to the same position as the heart along the anterior-posterior axis. The data represents the mean mitotic index of four different embryos per condition, with four sections analyzed per embryo. Error bars denote the standard deviation. (J) Tbx5 misexpression leads to an alteration in the timing and order of the cardiac differentiation program. RT-PCR analysis of the expression of Nkx2.5 as well as heart-specific isoforms of MHC, troponin and tropomyosin, and skeletal-muscle-specific genes, MyoD and muscle actin, throughout early and mid-gestation stages of development in control (‘C’) or Tbx5-injected (‘T’) stage-matched embryos. All samples are derived from a single batch of eggs and identical results were achieved in at least two independent sets of experiments for each marker. EF1-alpha was used as a control for all RT-PCR reactions.

Article Snippet: The antibodies against tropomyosin and cardiac troponin (developed by J.-C. Lin), and fibrillin (developed by C. D. Little) were obtained from the Developmental Studies Hybridoma Bank, developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA 52242.

Techniques: Injection, In Situ Hybridization, Expressing, Labeling, Standard Deviation, Control, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Marker

Journal: PLoS Biology

Article Title: A Ubiquitin Ligase Complex Regulates Caspase Activation During Sperm Differentiation in Drosophila

doi: 10.1371/journal.pbio.0050251

Figure Lengend Snippet: (A–H) Visualization of active drICE with anti-cleaved caspase-3 antibody (CM1; green) and axonemal tubulin polyglycylation with anti-glycylated tubulin monoclonal antibody (AXO 49; red). These figures are composed of a green layer only in the left panel, and green and red layers combined in the right panel. (A) Wild-type individualizing spermatids stain positively for active effector caspase and polyglycylated axonemal tubulin (white arrows pointing at cystic bulges [CBs] and red arrow pointing at a waste bag [WB]). Elongated spermatids from (B) homozygotes for the null cul3 mds1 allele or (C and D) transheterozygotes for cul3 mds1 and two different deficiencies that cover the cullin-3 gene, DF (2L)ED3 and DF (2L)Exel8034, respectively, stain for polyglycylation but not for active effector caspase. (E–G) Homozygote mutants for three hypomorphic cul3 Testis alleles, cul3 mds5 , cul3 mds3 , and cul3 mds4 , respectively, have spermatid individualization defects but still display some levels of active effector caspase expression. (H) However, the level of active effector caspase expression was dramatically reduced in spermatids from transheterozygote mutants for the null cul3 mds1 and either of the hypomorphic alleles, such as cul3 mds4 . All the figures are in the same magnification; scale bar 200 μm. (I) The diagram depicts a DEVDase activity assay for cul3 mds1 −/− testes. Caspase-3–like (DEVDase) activity is detected in wild-type testes and is blocked either after treatment with the caspase-3 inhibitor Z-VAD.fmk or in cul3 mds1 −/− testes. DEVDase activity, presented as relative luminescence units (RLUs), was determined on Ac-DEVD-pNA substrate in testis extracts made of 180 wild-type ( yw ) or cul3 mds1 −/− testes treated with Z-VAD or left untreated (DMSO). Readings were obtained every 2 min, and each time interval represents an average (mean ± SEM) of five readings. Note that the level of DEVDase activity in cul3 mds1 −/− testes is highly similar to the corresponding level in wild-type testes that were treated with Z-VAD. (J) A Western blot analysis for the assessment of the relative protein amounts used in (I). A portion of the testis extracts in (I) were used as controls to determine the relative amounts of total protein in each extract using the anti- β -Tubulin antibody.

Article Snippet: The tubes with 10 μl of the testes extracts were used for Western blot analysis to control for the protein amount in the samples by probing with anti- β -tubulin antibody (E7; 1:1000; Hybridoma Bank; http://dshb.biology.uiowa.edu/ ).

Techniques: Staining, Expressing, Activity Assay, Western Blot

Journal: PLoS Biology

Article Title: A Ubiquitin Ligase Complex Regulates Caspase Activation During Sperm Differentiation in Drosophila

doi: 10.1371/journal.pbio.0050251

Figure Lengend Snippet: (A) Diap1 protein levels were not affected in cul3 mds1 and klhl10 3 mutant testes, as assessed by Western blotting of protein extracts from dissected testes. Therefore, Diap1 does not appear to be a major target for the Cul3-based E3-ligase complex. β - tubulin protein levels served as loading control. (B andC) Co-IP experiment in S2 cells indicate that Klhl10 can bind to the BIR domain of dBruce. The immunoprecipitate (IP) is shown at the top, and pre-incubation of whole lysates are shown at the bottom (Input). Cell lysates were incubated with IgG beads which bind to Protein A (PrA). For Western blotting of IPs, (B) anti-dBruce antibody or (C) anti-HA antibody were used. (B) Cells were co-transfected with a dbruce “mini gene” (consisting of the first N-terminal 1,622 amino acids, including the BIR domain, and the last C-terminal 446 amino acids that contain the UBC domain) and (lane 1) PrA-klhl10 or (lane 2) PrA-GFP (see for details). (C) Cells were co-transfected with HA-tagged dBruce-BIR peptide containing the first N-terminal 387 amino acids of dBruce that includes the BIR domain region (amino acids 251–321). This motif is sufficient to bind to Klhl10 in S2 cells (see for details).

Article Snippet: The tubes with 10 μl of the testes extracts were used for Western blot analysis to control for the protein amount in the samples by probing with anti- β -tubulin antibody (E7; 1:1000; Hybridoma Bank; http://dshb.biology.uiowa.edu/ ).

Techniques: Mutagenesis, Western Blot, Control, Co-Immunoprecipitation Assay, Incubation, Transfection

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Dysregulated expression of primary cilium-associated genes in SARS-CoV-2 infected patient lung samples. A The Venn diagram presents genome-wide RNA sequencing results from patient lung samples, revealing differentially expressed genes (DEGs) associated with primary cilium. The DEGs were defined by log2 fold change > 1.5, with adjusted P < 0.05. P values were adjusted using the Benjamini–Hochberg correction. The analysis showing 3,972 DEGs in patient samples and 956 primary cilia-related genes in CiliaCarta, with 129 overlapping genes functionally linked to primary cilium. B The list of 129 ciliary DEGs identified in the SARS-CoV-2 patient lung samples. Gene ranked by the differently expressed level. Blue dots represent down-regulated genes, red dots represent up-regulated genes. Colour gradient and size of dots correlate with the degree of fold change. C Volcano plots of 129 DEGs in the patient lung samples overlap with the genes associated with ciliary functions. DEGs at -log10 > 2 and |log2 fold change|> 2 were indicated. (adjusted p values < 0.05, P values adjusted using the Benjamini–Hochberg correction) 61 up-regulated genes presented as red dots, 68 down-regulated genes presented as blue dots. Top 15 and two prominent ciliary genes ARL13B and ADCY3 are indicated. D Gene Ontology (GO) enrichment analysis for Biological Processes among ciliary DEGs from SARS-CoV-2 patient lung samples. Terms are ranked by their enrichment score, calculated using Gene Set Enrichment Analysis (GSEA). E Gene Set Enrichment Analysis (GSEA) reveals key signaling pathways associated with ciliary DEGs from SARS-CoV-2 patient lungs, ranked by enrichment score. F Ciliary DEGs from SARS-CoV-2 patient lung samples grouped by ciliary functional category. G Venn diagram and table show overlap between ACE2 interactors from BioGRID (979 proteins) and ciliary genes from CiliaCarta (956 genes) and Syscilia (303 genes). 56 and 14 overlapping genes were identified in each cilia database, respectively. Two well-studied ciliary components, namely ARL13B and TULP3 are identified in the intersections (highlighted yellow). H Gene Ontology (GO) enrichment analysis for Biological Processes among ciliary ACE2 interactors. Terms are ranked by their enrichment score, calculated using Gene Set Enrichment Analysis (GSEA)

Article Snippet: Cells were then incubated with anti-ACE2 antibody (#AF933, R&D systems) at room temperature for 2 h, followed by incubation with anti-ARL13B antibody (#N295B/66, Developmental Studies Hybridoma Bank) overnight at 4 °C.

Techniques: Expressing, Infection, Genome Wide, RNA Sequencing, Protein-Protein interactions, Functional Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: TULP3 colocalizes with ACE2 on the primary cilium axoneme and physically interacts with ACE2. A Representative immunofluorescence images show the accumulation and co-localization of TULP3 and ACE2 on the primary cilium of A549 cells. B Histogram illustrates co-localization profile of endogenous ACE2 and TULP3 expressions on ARL13B-positive cilia axoneme of A549. C Mander’s Coefficients analysis of ACE2 and TULP3 co-localization in cytoplasm versus primary cilia of A549 cell. The data represents quantifications from three independent experiments, with 10–20 data points measured in each experiment. Statistical analysis is performed using Paired Student’s t-Test, Two tailed. P values: ** < 0.01, *** < 0.001, **** < 0.0001. D Representative Western-blot images illustrate the co-IP experiment of ACE2 and TULP3 on ACE2-GFP overexpressing HEK293T cell line. IP: immunoprecipitation; IB: immunoblot. E Graph depicts densitometry analysis of TULP3 signal intensity in ACE2 and IgG immunoprecipitation groups. The data represents quantifications from three independent experiments. Statistical analysis is performed using Unpaired Student’s t-Test, Two tailed. P value * < 0.05. F Graph depicts densitometry analysis of ACE2 signal intensity in TULP3 and IgG immunoprecipitation groups. The data represents quantifications from three independent experiments. Statistical analysis is performed using Unpaired Student’s t-Test, Two tailed. P value * < 0.05. G Representative Western-blot images illustrate the co-IP experiment of ACE2 and ARL13B on ACE2-GFP overexpressing HEK293T cell line. IP: immunoprecipitation; IB: immunoblot. H TULP3–ACE2 interaction interface predicted from AlphaFold3 . TULP3 (light blue, residues 271–430) and ACE2 (light red, residues 749–773) are shown with key interface residues highlighted as sticks (blue for TULP3, red for ACE2). The TULP3 cargo-binding face (β8–β12, highlighted in marine) engages the membrane-proximal ACE2 segment. Yellow dashed lines represent top recurring contacts across models. ACE2 residue I761, involved in all major contacts, is labelled

Article Snippet: Cells were then incubated with anti-ACE2 antibody (#AF933, R&D systems) at room temperature for 2 h, followed by incubation with anti-ARL13B antibody (#N295B/66, Developmental Studies Hybridoma Bank) overnight at 4 °C.

Techniques: Immunofluorescence, Two Tailed Test, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation, Binding Assay, Membrane, Residue

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: TULP3 regulates ACE2 localization to the primary cilium. A Representative immunofluorescence images showing reduced ACE2 expression on the primary cilium in the shControl and TULP3 KD groups. B Graphs showing reduced ACE2 enrichment ratio in the (top) TULP3 and (bottom) ARL13B KD groups. ACE2 enrichment ratio was calculated as (ACE2 intensity per area on primary cilium)/(ACE2 intensity per area on whole cell). The data represents quantifications from three to four independent experiments, n = 14–17 measurements in each experiment. Statistical analysis is performed using (top) One-way ANOVA followed by Dunnett’s multiple comparisons and (bottom) Unpaired Student’s t-test. P values: * < 0.05, *** < 0.001, ns = not significant. C Representative immunofluorescence images show ciliary ACE2 expression on the primary cilium in TULP3 WT and TULP3 mut12 overexpression groups (pseudo-color magenta). D Graph shows ciliary ACE2 enrichment ratio in the TULP3 WT rescue and TULP3. mut12 overexpression groups. ACE2 enrichment ratio was calculated as (ACE2 intensity per area on primary cilium)/(ACE2 intensity per area on whole cell). The data represents quantifications from four independent experiments, n = 10–20 measurements in each experiment. Statistical analysis is performed using One-way ANOVA followed by Tukey’s multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001

Article Snippet: Cells were then incubated with anti-ACE2 antibody (#AF933, R&D systems) at room temperature for 2 h, followed by incubation with anti-ARL13B antibody (#N295B/66, Developmental Studies Hybridoma Bank) overnight at 4 °C.

Techniques: Immunofluorescence, Expressing, Over Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Attachment and accumulation of the viral spike protein on the ACE2-enriched primary cilium. A Representative immunofluorescence images show the negative control without exposure to spike protein. A ” Histogram illustrating the ciliary co-localization and expression profiles on of spike and ACE2 signals the negative control. B Representative immunofluorescence images show the accumulation and co-localization of viral recombinant spike protein on endogenously expressed ACE2 on the ARL13B. + primary cilium. B ” Histogram illustrating the co-localization and expression profile of three fluorescence channels reflecting viral spike protein, ARL13B and ACE2 respectively, on the primary cilium. C Graph depicts the spike expression intensity on ACE2-negative and ACE2-positive primary cilia. ACE2 pixel intensities per cilia area greater than 500 were defined as ACE2-positive primary cilia, whereas below 500 were counted as ACE2-negative primary cilia. The data represent quantifications from three independent experiments, with n = 10–20 for each group in each experiment. P values: ** < 0.01

Article Snippet: Cells were then incubated with anti-ACE2 antibody (#AF933, R&D systems) at room temperature for 2 h, followed by incubation with anti-ARL13B antibody (#N295B/66, Developmental Studies Hybridoma Bank) overnight at 4 °C.

Techniques: Immunofluorescence, Negative Control, Expressing, Recombinant, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Primary cilium and TULP3-dependent ciliary targeting of ACE2 in SARS-CoV-2 tropism

doi: 10.1186/s12964-025-02519-y

Figure Lengend Snippet: Genetic perturbation of primary cilia formation reduces SARS-CoV-2 infection in human host cells. A - B Representative Western blot images and the corresponding graphs show the knockdown efficiency of sh ARL13B and sh IFT88 in the ( A ) A549 and ( B ) RPE1 knockdown cells. Knockdown efficiency was quantified by normalizing the signal intensity of the target protein from each knockdown sample to its respective control group. The data represents quantifications from three independent experiments. Statistical analysis is performed using One-way ANOVA followed by Dunnett’s multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ns = not significant. Error bar: SD. C Representative immunofluorescence images show a reduced number of ciliated cells in the ARL13B KD and IFT88 KD groups in A549 (top panel) and RPE1 (bottom panel) cell lines. D - E Graphs depict the percentage of primary cilia-bearing (ciliated) cells in the ARL13B KD and IFT88 KD groups in ( D ) A549 and ( E ) RPE1 respectively. The data represents quantifications from five independent experiments. Statistical analysis is performed using One-way ANOVA followed by Dunnett’s multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ns = not significant. F - G Graphs depict flow cytometry analysis of the relative infection rate of different SARS-CoV-2-GFP pseudovirus variants upon perturbation of primary cilia in the ARL13B - and IFT88- knockdown human lung and retinal cells respectively. The percentage of infected cells was determined by flow cytometry analysis of GFP-positive infected cells. The data represents quantifications from three independent experiments. Each dot represents the mean value of an independent experiment. Statistical analysis is performed using Two-way ANOVA, Tukey's multiple comparisons test. P values: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. ns = not significant. Error bar: S.E.M. H Schematic diagram summarizes the findings, showing that the loss of ACE2-enriched cilia decreases SARS-CoV-2 viral uptake. Created with https://BioRender.com

Article Snippet: Cells were then incubated with anti-ACE2 antibody (#AF933, R&D systems) at room temperature for 2 h, followed by incubation with anti-ARL13B antibody (#N295B/66, Developmental Studies Hybridoma Bank) overnight at 4 °C.

Techniques: Infection, Western Blot, Knockdown, Control, Immunofluorescence, Flow Cytometry

Journal: NPJ Aging

Article Title: Mitochondrial respiratory chain deficiency is associated with an impaired skeletal muscle regenerative response and fibrosis in older men with HIV

doi: 10.1038/s41514-025-00273-6

Figure Lengend Snippet: A Representative fluorescence images of the three fibre types in men with HIV (n = 30) and men without HIV ( n = 15). Scale bar = 50 um. B Bar graph (mean + SD) showing the percentage of the three respective fibre types in men with HIV and men without HIV. C Representative fluorescence images of skeletal muscle cryo-sections stained with Pax7 (SCs) and DAPI (nuclei) in men with ( n = 30) and without HIV ( n = 15) individuals. Scale bar = 20 µm. D Graph (mean ± SD) showing the frequency of Pax7 + SCs per fibre. E Representative images of Masson’s trichrome-stained skeletal muscle sections in men with HIV ( n = 30) and men without HIV ( n = 15) for the determination of fibrosis. Scale bar = 100 µm. F Graph (mean ± SD) showing significantly greater levels of percentage fibrosis in men with HIV compared to men without HIV ( p < 0.0001). G Representative image of a H&E-stained skeletal muscle section showing degenerated (white arrow) and regenerated (yellow arrows demarking myofibres with central nuclei) myofibres. Scale bar = 50 µm. H Graph (median ± IQR) of the percentage regenerated fibres in men with HIV ( n = 30) and men without ( n = 15), with men with HIV displaying a significantly lower level of regenerated fibres ( p = 0.02). G Graph (median ± IQR) of the percentage degenerated fibres in men with HIV ( n = 30) and men without HIV ( n = 15).

Article Snippet: For the quantification of Pax7 + SC, 10 μm cryo-sections were air-dried for 1 h at RT before fixation in cold 4% PFA for 4 min. Next, sections were permeabilised for 10 min at RT with 0.2% Trion-x 100 before blocking with 5% NGS for 1 h at RT, followed by overnight incubation with Pax7 primary antibody (diluted in 10% NGS) (DSHB) at 4 °C.

Techniques: Fluorescence, Staining