Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Summary of the immunocytochemical analysis of CNHCs with antibodies against CD45, CD31, and CAIX

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (−UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX (A) . Clusters of CNHC-UMF and -BF without reactivity for CAIX antibodies ( B and C , respectively). A single CNHC-MF with positive cytoplasmic (D) and without staining for CAIX (E) . Single CAIX-negative CNHC-UMF and -BF ( F and G , respectively).

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques: Marker, Staining

Journal: PLoS ONE

Article Title: C9C5 positive mature oligodendrocytes are a source of Sonic Hedgehog in the mouse brain

doi: 10.1371/journal.pone.0229362

Figure Lengend Snippet: (A-B) Immunostaining of coronal brain sections from an adult mouse of the cerebral cortex and corpus callosum with the C9C5 antibody (yellow) and the oligodendroglial markers (A) CC1 (green) and Sox10 (red) or (B) CAII (green) and PDGFRα (red). White boxes (A, B) highlight magnifications of the cerebral cortex and the corpus callosum. (A) C9C5/CC1/Sox10 triple positive cells (white arrowhead) with stellate morphology are presented in merge and single channels with the nuclear marker DAPI. (B) C9C5 + (white arrowhead), CAII + (orange arrow), and PDGFRα + (white arrow) cells are presented in merge and single channels with the nuclear marker DAPI. Note that C9C5 positive cells are CAII and PDGFRα negative. Staining was replicated at least on three mice. Ctx, cerebral cortex; cc, corpus callosum.

Article Snippet: The primary antibodies were incubated overnight at 4°C: rabbit anti-SHHN (1/300, C9C5, #2207, Cell Signaling), mouse anti-GFAP (1/400, MAB360, Millipore), goat anti-Olig2 (1/400, AF2418, R&D Systems), mouse anti-S100β (1/500, S2532, Sigma), mouse anti-adenomatous polyposis coli (APC) (1/600, clone CC1, OP80, Millipore), chicken anti-βgalactosidase (1/200, ab9361, Abcam), goat anti-Sox10 (1/100, AF2864-SP, R&D Systems), rat anti-PDGFRα (1/300, 558774, BD Pharmingen), sheep anti-Carbonic Anhydrase II (CAII) (1/200, AHP206, BIORAD), rabbit anti-Iba1 (1/500, 019–19741, Fujifilm Wako), mouse anti-HuC/D (1/400, A-21271, Molecular Probes).

Techniques: Immunostaining, Marker, Negative Staining

Journal: PLoS ONE

Article Title: C9C5 positive mature oligodendrocytes are a source of Sonic Hedgehog in the mouse brain

doi: 10.1371/journal.pone.0229362

Figure Lengend Snippet: Quantification of C9C5 + cells in oligodendrocyte populations in the adult mouse brain.

Article Snippet: The primary antibodies were incubated overnight at 4°C: rabbit anti-SHHN (1/300, C9C5, #2207, Cell Signaling), mouse anti-GFAP (1/400, MAB360, Millipore), goat anti-Olig2 (1/400, AF2418, R&D Systems), mouse anti-S100β (1/500, S2532, Sigma), mouse anti-adenomatous polyposis coli (APC) (1/600, clone CC1, OP80, Millipore), chicken anti-βgalactosidase (1/200, ab9361, Abcam), goat anti-Sox10 (1/100, AF2864-SP, R&D Systems), rat anti-PDGFRα (1/300, 558774, BD Pharmingen), sheep anti-Carbonic Anhydrase II (CAII) (1/200, AHP206, BIORAD), rabbit anti-Iba1 (1/500, 019–19741, Fujifilm Wako), mouse anti-HuC/D (1/400, A-21271, Molecular Probes).

Techniques: Marker

Journal: Theranostics

Article Title: Hypoxia-induced TGFBI maintains glioma stem cells by stabilizing EphA2

doi: 10.7150/thno.95141

Figure Lengend Snippet: TGFBI is associated with the hypoxic microenvironment in human gliomas. (A) FISH staining of TGFBI in human GBM specimens (Ivy gap datasets). Scale bars: 2 mm; enlarged image: 100 μm. (B) Volcano map showing genes highly expressed in hypoxia region (pseudopalisade and microvascular proliferation regions). Each dot represents a gene. (C) Correlation between TGFBI and mRNA expression of hypoxia-related gene in the CCGA-GBM and TGGA-GBM datasets. CC, correlation coefficient; Dot size and color represent the correlation coefficient. (D) IF staining of TGFBI (green) and two hypoxia-associated markers, HIF1α (above, red) and CA9 (bottom, red), in human GBM specimens. Scale bars: 50 μm; enlarged image: 10 μm. (E) IHC staining demonstrating the association between TGFBI and HIF1α proteins in human gliomas. AOD, Average of density; n = 58.

Article Snippet: Primary antibodies: TGFBI (Abclonal, Cat#A11222, for IB, 1:1000; for IP, 5ug; Proteintech, Cat#10188-AP, for IHC, 1:100; for IF, 1:100), CD133 (Affinity, Cat#BF0403, for IF, 1:100), SOX2 (Proteintech, Cat#66411-1-Ig, for IB, 1:1000; Santa Cruz, Cat#365823, for IF, 1:50; for IHC, 1:50), HIF1α (Proteintech, Cat#20960-1-AP, for IB, 1:1000; for IF, 1:50; for IHC 1:50), carbonic anhydrase 9 (CA9, NOVUS, Cat#NB100-417, for IF, 1:50), OLIG2 (Proteintech, Cat#66513-1-Ig, for IB, 1:1000; for IF, 1:200), Ki67 (Proteintech, Cat#27309-1-AP, for IF, 1:200), EphA2 (Abclonal, Cat#A7183, for IB,1:1000; Santa Cruz, Cat#398832, for IF, 1:50; for IHC, 1:50; for IP, 5ug), Akt (Proteintech, Cat#60203-1-Ig, for IB, 1:5000), phospho-Akt (Ser473) (Proteintech, Cat#66444-1-Ig, for IB, 1:2000; Cell Signaling Technology, Cat#4060, for IF, 1:100; for IHC, 1:50), c-MYC (Proteintech, Cat#67447-1-Ig, for IB, 1:5000; for IF,1:100; for IHC, 1:100), phospho-EphA2 (Y588) (Proteintech, Cat#30263-1-AP, for IB, 1:1000), Integrin αV (Proteintech, Cat#27096-1-AP, for IB, 1:1000), Integrin β5 (CST, Cat#3629, for IB, 1:1000), Notch1 (Cell Signaling Technology, Cat#3608, for IB, 1:1000).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: bioRxiv

Article Title: Hippocampal cell- and circuit-specific differences in mitochondrial form and function

doi: 10.64898/2025.12.16.694759

Figure Lengend Snippet: A.Representative coronal image of the hippocampus showing CA2 neurons (labeled with RGS14) and sparse, cre-dependent labeling of mitochondria (Mito-GFP) B.Representative high magnification images of Mito-GFP and RFP-labeled secondary dendrites in CA1 SR (i) and SLM (ii). C.Representative high magnification images of Mito-GFP and RFP-labeled secondary dendrites in CA2 SR (i) and SLM (ii). MitoTag-GFP images are scaled to the same fluorescence range. RFP images are scaled to visualization. D.Histogram showing the percent cumulative frequency of non-normalized mitochondria areas in CA1 and CA2 SR and SLM. CA1 SR n = 1866 mitochondria from 111 dendrites, SLM n = 2387 mitochondria from 122 dendrites; CA2 SR n = 1653 mitochondria from 96 dendrites, CA2 SLM n = 2159 mitochondria from 119 dendrites from 8 mice. E.Bar plot of average mitochondrial areas per animal (N= 8 mice). Data are normalized to each animal average. (paired two-way ANOVA, main effect of subregion F(1, 7) = 33.32, p=0.0007 and main effect of layer F(1, 7) = 86.18, p<0.0001, Fisher’s LSD posthoc tests are shown on the plot) F.Bar plot of average dendrite areas per animal (N= 8 mice). Data are normalized to each animal average. (paired two-way ANOVA, no effect of subregion F (1, 7) = 4.069, p=0.0835, no effect of layer F(1, 7) = 0.9857 p=0.3539) G.Bar plot of percent area of the dendrite occupied by mitochondria. (paired two-way ANOVA, main effect of subregion F(1, 7) = 16.11, p=0.0051, and layer F(1, 7) = 39.96, p=0.0004, Fisher’s LSD posthoc tests are shown on the plot)*p <0.05; **p<0.01; ***p<0.001. Scale bars = 100 µm (A), 2 µm (C). Error bars are average ± SEM.

Article Snippet: Sections were immunostained with mouse-anti-RGS14 (1:500, NeuroMab cat# 75-170, RRID: AB_2877352) as a CA2 marker as well as rabbit-anti-OPA1 (1:200, Proteintech cat# 27733-1-AP, RRID: AB_2810292) or rabbit-anti-MFF (1:500, Proteintech cat# 17090-1-AP, RRID: AB_2142463).

Techniques: Labeling, Fluorescence

Journal: bioRxiv

Article Title: Hippocampal cell- and circuit-specific differences in mitochondrial form and function

doi: 10.64898/2025.12.16.694759

Figure Lengend Snippet: A.Representative coronal tile image of OPA1 and CA2 marker RGS14 in the hippocampus. Arrowheads denote CA2 cell body borders. B.The same image as in A with only OPA1. C.Bar plot of OPA1 fluorescence intensity in CA1 and CA2 SP, SR and SLM normalized to the animal average. (Paired two-way ANOVA, main effect of subregion (F(1,7) =6.623, p=0.0368), layer (F(2,14)=52.02, p<0.0001), and interaction effect of subregion x layer (F(2,14) =86.69, p<0.0001, N = 8 mice, Tukey’s posthoc tests are shown on the plot). D.Representative coronal tile image of MFF and CA2 marker RGS14 in the hippocampus. Arrowheads denote CA2 cell body borders. E.The same image as in D with only MFF. F.Quantification of MFF fluorescence intensity in CA1 and CA2 SP, SR and SLM normalized to the animal average. (Paired two-way ANOVA, main effect of subregion (F(2, 14) = 306.9, p<0.0001) with an interaction effect of subregion x layer (F(2, 14) = 32.21, p<0.0001, N = 8 mice, Tukey’s posthoc tests are shown on the plot). ***p<0.001 ; ****p<0.0001. Scale bar = 100 µm. Error bars are average ± SEM.

Article Snippet: Sections were immunostained with mouse-anti-RGS14 (1:500, NeuroMab cat# 75-170, RRID: AB_2877352) as a CA2 marker as well as rabbit-anti-OPA1 (1:200, Proteintech cat# 27733-1-AP, RRID: AB_2810292) or rabbit-anti-MFF (1:500, Proteintech cat# 17090-1-AP, RRID: AB_2142463).

Techniques: Marker, Fluorescence

Journal: bioRxiv

Article Title: Hippocampal cell- and circuit-specific differences in mitochondrial form and function

doi: 10.64898/2025.12.16.694759

Figure Lengend Snippet: A.Representative coronal live slice images of CA1 mitochondrial calcium pre (i) and post KCl treatment (ii). B.Normalized average mito-RGECO intensity line plots in CA1 layers relative to distance from stratum oriens (SO, 0.0). The thin lines represent the average trace per mouse normalized by the average intensity within each slice. Multiple slices from the same mouse were averaged to produce a single line per mouse. The thicker line is the average of all animals. (N=6 mice, 11 slices). C.Representative horizontal live slice images of CA2 mitochondrial calcium at baseline (i) and post KCl treatment (ii). D.Normalized average mito-RGECO intensity line plots in CA2 layers as in B. (N=9 mice, 12 slices). E.Quantification of KCl-induced change in mitochondrial calcium fluorescence normalized to baseline (ΔF/F) plotted as an average per animal (N=6-9 mice, 1-3 slices per mouse, 11 CA1 slices, 12 CA2 slices). No significant differences in subregion, layer, or interactions (two-way ANOVA). F.The ratio of mitochondrial calcium signals in SLM to SR at baseline and post KCl treatment (two-way ANOVA, main effect of subregion, F (1,13) = 8.244, p=0.0136, Fisher’s LSD posthoc tests are shown on the plot). G.Representative coronal live slice images of CA1 GCAMP6f cytosolic calcium levels pre (i) and post KCl (ii). The same slice is shown as in A. H.Normalized average GCaMP6f intensity line plots in CA1 layers as in B. I.Representative horizontal live slice images of CA2 GCAMP6f cytosolic calcium levels pre (i) and post KCl (ii). The same slice is shown as in C. J.Normalized average GCAMP6f intensity line plots in CA2 layers as in B. Note the lack of a peak in SLM relative to SR as seen in D. *p<0.05 p**<0.01. Scale = 100 µm. Error bars are average ± SEM. The brightness of images in A, C, G & I are optimized per slice.

Article Snippet: Sections were immunostained with mouse-anti-RGS14 (1:500, NeuroMab cat# 75-170, RRID: AB_2877352) as a CA2 marker as well as rabbit-anti-OPA1 (1:200, Proteintech cat# 27733-1-AP, RRID: AB_2810292) or rabbit-anti-MFF (1:500, Proteintech cat# 17090-1-AP, RRID: AB_2142463).

Techniques: Fluorescence

Journal: bioRxiv

Article Title: Hippocampal cell- and circuit-specific differences in mitochondrial form and function

doi: 10.64898/2025.12.16.694759

Figure Lengend Snippet: A.Representative tile image of mito-RGECO (i) and CA2 marker RGS14 (ii) in a perfusion fixed, coronal section of the hippocampus (dashed line denotes the border of SR and SLM). B.Representative high magnification image of RGS14 labeled CA2 neurons (cyan) expressing mito-RGECO (magenta) and sparse, cre-dependent labeling of MitoTag-GFP (green). Insets of a CA2 cell soma and proximal dendrites. Note the mito-RGECO signal is localized to the mitochondrial matrix surrounded by the GFP-labeled outer membrane. C.Representative images of mito-RGECO in CA1 (i) and CA2 (ii). Insets show mito-RGECO in distal dendrites. D.Normalized average mito-RGECO intensity line plots across CA1 and CA2 layers relative to distance from stratum oriens (SO). Data points are normalized by the average intensity within a subregion per animal. Thus, CA1 and CA2 data should not be directly compared, but rather the layer differences within each subregion. The data shown are animal averages ± SEM. (CA1 N=5 mice, CA2 N=7 mice). E.Average ratio of SR to SLM mito-RGECO fluorescence intensities. (unpaired two tailed t-test, CA1 N=5 mice, CA2 N=7 mice) ***p<0.0001. Scale bars = 100 µm (A, C), 20 µm (B), 5 µm (B, C insets). Error bars are average ± SEM.

Article Snippet: Sections were immunostained with mouse-anti-RGS14 (1:500, NeuroMab cat# 75-170, RRID: AB_2877352) as a CA2 marker as well as rabbit-anti-OPA1 (1:200, Proteintech cat# 27733-1-AP, RRID: AB_2810292) or rabbit-anti-MFF (1:500, Proteintech cat# 17090-1-AP, RRID: AB_2142463).

Techniques: Marker, Labeling, Expressing, Membrane, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Hippocampal cell- and circuit-specific differences in mitochondrial form and function

doi: 10.64898/2025.12.16.694759

Figure Lengend Snippet: A.Post-fixed CA1 coronal slice stained for GFP (GCaMP6f, i), RFP (mito-RGECO, ii), and CA1 marker CALB1 (iii). Zoomed inset of SP (iv) to show mito-RGECO expression in CALB1 expressing CA1 neurons. B.Post-fixed CA2 horizontal slice stained for GFP (GCaMP6f, i), RFP (mito-RGECO, ii), and CA2 marker NECAB2 (iii). Zoomed inset of SP (iv) to show mito-RGECO expression in NECAB2 expressing CA2 neurons. C.Representative tile image of sparse hSyn1-cre driven tdtomato labeling of CA1 and CA2 neurons in a coronal section. CA2 neurons are labeled with PCP4. D.Higher magnification of CA1 to show dendritic banding in SLM. Nuclei are labeled with DAPI. E.Higher magnification of CA2 to show dendrites are out of plane. Nuclei are labeled with DAPI. Scale bars = (Ai) 100 µm, (Aiv) 50 µm, (C) 100 µm, (D) 100 µm

Article Snippet: Sections were immunostained with mouse-anti-RGS14 (1:500, NeuroMab cat# 75-170, RRID: AB_2877352) as a CA2 marker as well as rabbit-anti-OPA1 (1:200, Proteintech cat# 27733-1-AP, RRID: AB_2810292) or rabbit-anti-MFF (1:500, Proteintech cat# 17090-1-AP, RRID: AB_2142463).

Techniques: Staining, Marker, Expressing, Labeling

Journal: Medical Oncology (Northwood, London, England)

Article Title: Expression of HIF-1α and CAIX in nasopharyngeal carcinoma and their correlation with patients’ prognosis

doi: 10.1007/s12032-014-0304-1

Figure Lengend Snippet: Expression of HIF-1α and CAIX by immunohistochemistry in NPC tissues (original magnification ×400). a Nuclear membrane positive staining of HIF-1α in NPC tissue. b Plasmalemma and cytoplasmic positive staining of CAIX in NPC tissue

Article Snippet: Each section was incubated with protein blocker for 15 min at room temperature followed by mouse anti-human HIF-1α antibody(1:100, Novus Biologicals, Colorado, USA) and rabbit anti-human CAIX antibody (1:400, Novus Biologicals, Colorado, USA)incubation at 4 °C overnight.

Techniques: Expressing, Immunohistochemistry, Membrane, Staining

Journal: Medical Oncology (Northwood, London, England)

Article Title: Expression of HIF-1α and CAIX in nasopharyngeal carcinoma and their correlation with patients’ prognosis

doi: 10.1007/s12032-014-0304-1

Figure Lengend Snippet: The Kaplan–Meier curves for NPC patients with both positive and negative expressions of HIF-1α and CAIX: a overall survival, b progression-free survival, c loco-regional relapse-free survival, d distant metastasis-free survival

Article Snippet: Each section was incubated with protein blocker for 15 min at room temperature followed by mouse anti-human HIF-1α antibody(1:100, Novus Biologicals, Colorado, USA) and rabbit anti-human CAIX antibody (1:400, Novus Biologicals, Colorado, USA)incubation at 4 °C overnight.

Techniques: