rabbit anti phox2b antibody  (Proteintech)

Journal: Animal Cells and Systems

Article Title: Systematic evaluation of AAV administration strategies for targeting Phox2b + primary visceral sensory neurons

doi: 10.1080/19768354.2026.2749248

Figure Lengend Snippet: Systemic administration of AAV-PHP.S-Phox2b-Cre at P0. (A) Schematic for subcutaneous injection of AAV-PHP.S-Phox2b-Cre (1.68 × 10¹¹ vg/mouse) into P0 neonates (<1.5 g). (B–H) Representative confocal images showing tdTomato fluorescence in the indicated organs. (B) Tongue. Anti-PGP9.5 (green) and tdTomato (red). (C) Tongues. Anti-Krt8 (green) and tdTomato (red). (D) Small intestine. Anti-PGP9.5 (green) and tdTomato (red). (E) NPG. Anti-NeuN (green) and tdTomato (red). (F) Hindbrain showing the rostral (bregma, −7.15 mm) and caudal (bregma, −7.55 mm) NTS. Anti- NeuN (green) and tdTomato (red). (G) Choroid plexus sections. Anti-Phox2b (green) and tdTomato (red). (H) Forebrain (bregma, 1.42 mm); dashed lines delineate the cortex and subcortex, with a magnified view of ectopic tdTomato expression in the cortical region. Scale bars: 50 μm (B, C), 100 μm (D, E), and 500 μm (F–H). n = 3 mice.

Article Snippet: The following primary antibodies were used in this study: anti-PGP9.5 (1:400, GTX109637, GeneTex), anti-tdTomato (1:200, AB8181-200, Sicgen), anti-Krt8 (1:1000, ab53280, Abcam), anti-NeuN (1:1000, ab177487, Abcam), and anti-Phox2b (1:200, sc-376997, Santa Cruz).

Techniques: Injection, Confocal, Fluorescence, Expressing

Journal: bioRxiv

Article Title: ALK R1275Q mutation drives expansion of SCP-like cells during sympathoadrenal commitment and primes neuroblastoma initiation

doi: 10.64898/2026.01.27.701690

Figure Lengend Snippet: a, Schematic representation of the differentiation protocol. The first row shows the transition from iPSC reprogramming to tNCC and subsequently to sympathoadrenal (SA) differentiation. Dashed lines of different colors indicate time points subjected to single-cell RNA sequencing, consistently applied throughout this study. The second row defines the project time line, with the tNCC stage designated as day 0. The third row lists bulk (B) or single-cell (S) RNA-seq performed at each collection point, and the fourth row indicates the names used for subsequent analyses. Rows five and six show the plate-coating substrates and growth factors used in culture. Representative merged immunofluorescence images (right) show iPSCs (OCT4, SSEA4), tNCCs (HOXC9, SOX10), sympathoadrenal progenitors (SAP; PHOX2B, KI67), and sympathoadrenal mature cells (SAM, day 12; PRPH). Nuclei were counterstained with DAPI. Scale bar, 100 µm. Corresponding single-channel images are available in the Supplementary Figures. b, Expression of selected HOX family genes was analyzed to assess anterior–posterior identity. Color-coded cranial (pink), trunk (green), and caudal markers (blue). The bubble plot illustrates HOX gene expression levels across collection time points (log₂[TPM+1]). c, Bubble plot showing expression of selected markers of NCC-to-SA lineage differentiation, based on published datasets of human adrenal medulla development. Values are shown as average log₂ (TPM+1). d, UMAP visualization of single-cell RNA-seq data from Ctrl cell differentiation. Cells were assigned to ten transcriptional clusters, indicated by colors and annotated (as detailed in the Methods section) by collection time point (upper left), individual identity (lower left), and cell-cycle phase (upper right), and SYM-MES score (lower right). The circular plot surrounding the UMAP shows cluster composition by time point and cell-cycle status, arranged clockwise by time point numbers indicate the log₁₀-scaled angular positions of ordered cell groups along the circular layout (middle).

Article Snippet: Primary antibodies (overnight, 4 °C): Ki67 (1:100, Abcam ab16667), PHOX2B (1:100, Abclonal A21990).

Techniques: RNA Sequencing, Immunofluorescence, Expressing, Gene Expression, Cell Differentiation

Journal: bioRxiv

Article Title: ALK R1275Q mutation drives expansion of SCP-like cells during sympathoadrenal commitment and primes neuroblastoma initiation

doi: 10.64898/2026.01.27.701690

Figure Lengend Snippet: a, Representative immunofluorescence images showing tNCC expression of trunk marker HOXC9 and neural crest marker SOX10. Scale bars, 100 µm. The image marked with an asterisk is also shown in . The separated channels of the immunofluorescence are shown in . b, RT–qPCR analysis of marker expression across differentiation stages. Markers include pluripotency ( OCT4 , NANOG ), neural crest ( PAX3 , SNAI2 , SOX10 , TFAP2A ), trunk ( HOXC9 , CDX2 ), and cranial ( OTX2 ). Each dot represents a biological replicate (n=6), and lines connect paired samples. Expression of each gene was normalized to Ctrl7 iPSCs. Data are shown as mean ± s.d. Statistical significance was determined using a paired two-tailed Student’s t -test. tNCC vs iPSC in OCT4 , NANOG , PAX3 , SNAI2 , SOX10 , TFAP2A and HOXC9 , P < 0.0001 = ****; in CDX2 , P = 0.002 = **; in OTX2 , P = 0.0015 = **. c, Representative immunofluorescence images showing SAP expression of the sympathoadrenal lineage marker PHOX2B and the proliferation marker KI67. Scale bars, 100 µm. The image marked with an asterisk is also shown in . The separated channels of the immunofluorescence are shown in . d, Representative immunofluorescence images showing SAM expression of the peripheral nervous system (PNS) marker PRPH (top), the proliferation marker KI67, the mature sympathoadrenal marker ISL1/2 (middle), the catecholaminergic marker TH, and the neuronal marker TUBB3 (bottom). Scale bars, 100 µm. The image marked with an asterisk is also shown in . The separated channels of the immunofluorescence are shown in - .

Article Snippet: Primary antibodies (overnight, 4 °C): Ki67 (1:100, Abcam ab16667), PHOX2B (1:100, Abclonal A21990).

Techniques: Immunofluorescence, Expressing, Marker, Quantitative RT-PCR, Two Tailed Test

Journal: bioRxiv

Article Title: ALK R1275Q mutation drives expansion of SCP-like cells during sympathoadrenal commitment and primes neuroblastoma initiation

doi: 10.64898/2026.01.27.701690

Figure Lengend Snippet: a, UMAP of all samples (as in ), colored by ALK expression. b, Violin plots of ALK expression (top) and ALK activity scores based on Reactome pathways (bottom) across time points in Ctrl and patient lines (computed as detailed in Methods). Comparisons were performed using the Wilcoxon test. ALK expression, Ctrl vs NB in SAP, D8, and D12, P < 0.0001 = ****; ALK Reactome score, Ctrl and NB in tNCC, D8, and D12, P < 0.0001 = ****, in SAP, P = 0.036 = *. Black triangles indicate mean values. c, Immunoblot analysis of protein lysates collected at NCC, SAP, day 8 (D8), and day 12 (D12) from two Ctrl lines and two patient-derived lines. Total and phosphorylated ALK, AKT, and ERK1/2 were probed. GAPDH was used as a loading control. P.C., positive control: NB-1 neuroblastoma cell line (CVCL_1440) treated with ALKAL2 . d, Quantification of EdU+ cells at the SAP stage after Lorlatinib treatment of both Ctrl (Ctrl 7, Ctrl10, and Ctrl14 in blue) and patient-derived (NB1, NB2, and NB4 in red) lines. Quantification of EdU⁺ cell percentages is shown as mean ± s.d. from independent biological replicates. Statistical significance was assessed by one-way ANOVA followed by Šídák’s multiple comparisons test. In DMSO, Ctrl vs NB, P < 0.0001 = ****; in NB, lorlatinib (20nM) vs DMSO, P < 0.0001 = ****; Lorlatinib (30nM) vs DMSO, P < 0.0001 = ****. e, Schematic of in vivo experimental design. iPSCs were stably transfected with a vector encoding constitutively expressed nuclear mCherry and doxycycline-inducible MYCN . Differentiated tNCCs were injected subcutaneously or orthotopically into the adrenal gland. MYCN was activated by mice receiving doxycycline (200 μg/ml) in drinking water. f, Kaplan–Meier survival curves from the subcutaneous injection model. Solid line, mice treated with doxycycline; dashed line, untreated controls. Significance was assessed by Cox proportional hazards test. g, Table summarizing results of the orthotopic injection experiment. Tumor size is reported as the longest axis of the tumor. h, Histological analysis of tumors from orthotopically injected mice. Gross tumor morphology is shown (with ruler indicating size, top). Representative sections were stained for PHOX2B, KI67, and hematoxylin and eosin (H&E) in the same tumor region. Scale bar, 100 µm.

Article Snippet: Primary antibodies (overnight, 4 °C): Ki67 (1:100, Abcam ab16667), PHOX2B (1:100, Abclonal A21990).

Techniques: Expressing, Activity Assay, Western Blot, Derivative Assay, Control, Positive Control, In Vivo, Stable Transfection, Transfection, Plasmid Preparation, Injection, Staining