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: Cells, Tissues, Organs

Article Title: Carbonic Anhydrase IV Deficiency Causes Intrauterine Embryonic Loss in Mice

doi: 10.1159/000544000

Figure Lengend Snippet: CAIV immunostaining of reproductive organs in nonpregnant mice. a–d Ovary. No immunostaining is visible in the mullerian epithelium (ME in a ), ovarian stroma (S in a ), primordial follicles (arrow in a ), primary follicles (* in b ), secondary follicles (SF in b ), tertiary follicles ( c ), and corpora lutea ( d ). e , f Fallopian tube. Neither in the ampulla ( e ) nor in the isthmus ( f ) is CAIV immunostaining observed in the epithelium (Ep) and musculature (M). In the uterus ( g ), endometrial capillaries were CAIV immunoreactive (insert in g ), while no staining is visible in the endometrial epithelium (Ep) and the endometrial stromal cells (S). h Vagina. In the vaginal tissue, no CAIV-immunoreactive cells are observed in the epithelium (Ep) or in the stroma (S). i In kidney as positive control, a strong CAIV signal is visible in tubular epithelium while the negative control of the endometrium ( j ) shows no immunostaining (counterstaining with hematoxylin). Bar, 50 μm.

Article Snippet: After rinsing in distilled water, slices were blocked with PBS-glucose-oxidase buffer and incubated with a polyclonal goat IgG antibody against CAIV (AF2414; R&D Systems, Minneapolis, MN, USA) (1:100 in PBS/5% BSA) overnight at 4°C, followed by incubation with biotinylated anti-goat IgG (1:200 in PBS/5% BSA) for 30 min at room temperature.

Techniques: Immunostaining, Staining, Positive Control, Negative Control

Journal: Cells, Tissues, Organs

Article Title: Carbonic Anhydrase IV Deficiency Causes Intrauterine Embryonic Loss in Mice

doi: 10.1159/000544000

Figure Lengend Snippet: CAIV immunostaining of mouse embryos at developmental stages 4.5 ( a ), 5.5 ( b ), 6.5 ( c ), and 7.5 dpc ( d ). CAIV immunoreactivity is visible in the blastocyst (B), the visceral endoderm (VE), and the embryonic ectoderm (IE). Corresponding negative controls ( a ′, b ′, c ′, d ′) show no immunostaining. Slides are counterstained with hematoxylin. Bar, 50 μm in a , b , 100 μm in c , d , 100 μm in a ′, b ′, and 160 μm in c ′, d ′.

Article Snippet: After rinsing in distilled water, slices were blocked with PBS-glucose-oxidase buffer and incubated with a polyclonal goat IgG antibody against CAIV (AF2414; R&D Systems, Minneapolis, MN, USA) (1:100 in PBS/5% BSA) overnight at 4°C, followed by incubation with biotinylated anti-goat IgG (1:200 in PBS/5% BSA) for 30 min at room temperature.

Techniques: Immunostaining

Journal: Cells, Tissues, Organs

Article Title: Carbonic Anhydrase IV Deficiency Causes Intrauterine Embryonic Loss in Mice

doi: 10.1159/000544000

Figure Lengend Snippet: CAIV immunostaining of a representative mouse embryo at 8.5 dpc in overview ( a ) and in higher magnification ( b , c ). A strong immunoreactivity is visible in the amniotic membrane (AM), the yolk sac epithelium (Y), trophoblast giant cells (TGC), the endometrial epithelium (EE), and the developing GT of the embryo (E). Corresponding negative controls ( a ′, b ′, c ′) show no immunostaining. Slides are counterstained with hematoxylin. Bar indicates 200 μm in a and 50 μm in b , c .

Article Snippet: After rinsing in distilled water, slices were blocked with PBS-glucose-oxidase buffer and incubated with a polyclonal goat IgG antibody against CAIV (AF2414; R&D Systems, Minneapolis, MN, USA) (1:100 in PBS/5% BSA) overnight at 4°C, followed by incubation with biotinylated anti-goat IgG (1:200 in PBS/5% BSA) for 30 min at room temperature.

Techniques: Immunostaining, Membrane

Journal: Cells, Tissues, Organs

Article Title: Carbonic Anhydrase IV Deficiency Causes Intrauterine Embryonic Loss in Mice

doi: 10.1159/000544000

Figure Lengend Snippet: CAIV immunostaining of mouse embryos at 9.5 dpc in overview ( a , b ) and in higher magnification ( c–h ). Within the developing embryo ( c , d ), CAIV immunostaining is visible in the gut tube (GT), the floor plate of the neural tube (FP), and the notochord (N). In placenta and extraembryonic tissues ( e–h ), the labyrinthine compartment (LT), the endometrial epithelium (EE), the trophoblast giant cells (TGC), and the yolk sac epithelium (Y) show immunoreactivity. The corresponding negative control ( b ) shows no immunostaining (counterstaining with hematoxylin). Bar = 500 μm in a , b and 50 μm in c–h .

Article Snippet: After rinsing in distilled water, slices were blocked with PBS-glucose-oxidase buffer and incubated with a polyclonal goat IgG antibody against CAIV (AF2414; R&D Systems, Minneapolis, MN, USA) (1:100 in PBS/5% BSA) overnight at 4°C, followed by incubation with biotinylated anti-goat IgG (1:200 in PBS/5% BSA) for 30 min at room temperature.

Techniques: Immunostaining, Negative Control

Journal: Cells, Tissues, Organs

Article Title: Carbonic Anhydrase IV Deficiency Causes Intrauterine Embryonic Loss in Mice

doi: 10.1159/000544000

Figure Lengend Snippet: Litter size and sex distribution in heterozygous and CAIV knockout mating. a Mating of CAIV knockout mice ( n = 17 litters) results in a significant reduction of litter size compared to heterozygous mating ( n = 31 litters). b Homozygous CAIV knockout mating results in a non-mendalian sex distribution of the 54 CAIV knockout pubs analyzed (74% male and 26% female instead of 50% each). c In heterozygous mating, the 164 pups analyzed show an enhanced number of wild-type (31% instead of 25% expected) and heterozygous genotype (62% instead of 50% expected), whereas the number of CAIV knockout pups is significantly reduced (7% instead of 25% expected). d A non-mendelian sex distribution is also observed in the 12 CAIV knockout pups (shown in c ) resulting from heterozygous mating (67% male and 33% female instead of 50% each), while there is a slight preponderance of female pups in both the 50 resulting wild-type (36% male vs. 64% female) and the 102 resulting CAIV heterozygous pups (46% male vs. 54% female). e On 9.5 dpc, number of implantation chambers is similar in heterozygous ( n = 3) and CAIV knockout ( n = 5) mating. The sex distribution of embryos is slightly shifted to female in both experimental groups, showing a higher percentage of female embryos in heterozygous mating in total ( n = 26; 40% male vs. 60% female) as well as in CAIV knockout embryos resulting from heterozygous mating (40% male vs. 60% female). f Additionally, CAIV knockout mating results in a higher percentage of female embryos on day 9.5 of pregnancy ( n = 21; 43% male vs. 57% female).

Article Snippet: After rinsing in distilled water, slices were blocked with PBS-glucose-oxidase buffer and incubated with a polyclonal goat IgG antibody against CAIV (AF2414; R&D Systems, Minneapolis, MN, USA) (1:100 in PBS/5% BSA) overnight at 4°C, followed by incubation with biotinylated anti-goat IgG (1:200 in PBS/5% BSA) for 30 min at room temperature.

Techniques: Knock-Out

Journal: International Journal of Biological Sciences

Article Title: Inhibition of GRK2-PDE4D Axis Suppresses Fibroblast-Like Synoviocytes Hyperplasia and Alleviates Experimental Arthritis

doi: 10.7150/ijbs.100176

Figure Lengend Snippet: Grk2 -deficient mice relieved symptoms in CAIA. (a) Male Grk2 +/+ and Grk2 +/- mice were used to establish a CAIA mouse model as indicated. (b) Swollen joint count and (c) arthritis index of mice were recorded from the 0th day to the 12th day after intraperitoneal injection of mouse monoclonal anti-collagen-II 5-clone antibody cocktail. n = 5. (d) Representative photographs of paws (upper) and H&E staining (lower) of the ankle joint sections from different groups. Histopathologic changes include synovial hyperplasia (red arrowhead), pannus (blue arrowhead), infiltrating inflammatory cells (green arrowhead), and cartilage destruction (yellow arrowhead). (e) Histopathological score of inflammation, cartilage erosion, pannus formation, and synoviocytes proliferation of Grk2 +/+ and Grk2 +/- CAIA mice. n = 5. (f) Representative double-staining IF of PDE4D and GRK2 from the synovial tissues of normal and CAIA mice. (g) Quantification of vimentin and (h) PDE4D MFI in the synovial tissues of normal and CAIA mice. n = 5. (i) The levels of TNF-α in mice serum were measured by ELISA. n = 5. (j) Representative p-ERK and P65 IF staining in the synovium of CAIA mice. Data are presented as mean ± SD from at least five independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: 2) Mouse collagen antibody-induced arthritis (CAIA) model: For the induction of the mouse CAIA model, male Grk2 +/+ and Grk2 +/- mice (10-12-weeks-old) were intraperitoneally injected with 4 mg of anti-collagen-II 5-clone antibody cocktail (53100, Chondrex, Redmond, WA, USA) on day 0, followed by a subsequent intraperitoneal injection of 40 mg lipopolysaccharide on day 3, as previously described .

Techniques: Injection, Staining, Double Staining, Enzyme-linked Immunosorbent Assay

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: Acta Pharmaceutica Sinica. B

Article Title: An immunostimulant nanomedicine enhances radioimmunotherapy by remodeling the tumor immunosuppressive landscape after radiotherapy

doi: 10.1016/j.apsb.2025.11.012

Figure Lengend Snippet: Immunosuppression after radiotherapy promotes breast cancer progression. Tumor growth curves (A) and weights (B) of mice before and after RT ( n = 6). (C) Survival curve of mice after different treatments ( n = 6). Tumor growth curves (D) and weights (E) of mice treated with RT and RT plus α -PD-L1 ( n = 6). (F) Survival curve of mice after different treatments ( n = 6). (G) Cluster analysis of differential expression genes between untreated and RT-treated tumors 48 h post-RT ( n = 3). (H) Gene Ontology (GO) analysis of tumor tissues before and after RT (select the top 10 for each item). (I) Heat map of differentially expressed genes related to apoptosis and immune suppression ( n = 3). (J) Immunofluorescence staining images of tumor tissue in saline and RT groups (scale bar = 20 μm). (K) Quantitative analysis of tumor-infiltrating CD45 + cells ( n = 6). Representative flow cytometry images (M) and quantitative analysis (L) of tumor-infiltrating MDSCs ( n = 6). (N) The TUNEL staining of tumor section (scale: 25 μm). (O) Adenosine content detection in tumor tissue ( n = 6). Data are presented as mean ± SD. ∗∗∗∗ P < 0.0001 determined by Student’s t-test.

Article Snippet: InVivo MAb anti-mouse PD-L1 was purchased from Bioxcell (USA).

Techniques: Quantitative Proteomics, Immunofluorescence, Staining, Saline, Flow Cytometry, TUNEL Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: An immunostimulant nanomedicine enhances radioimmunotherapy by remodeling the tumor immunosuppressive landscape after radiotherapy

doi: 10.1016/j.apsb.2025.11.012

Figure Lengend Snippet: FD@ATRA-enhanced radiotherapy combined with α -PD-L1 to enhance the efficacy of large-volume tumors. (A) Schematic diagram of the therapeutic process for evaluating the anti-tumor effects in a bilateral 4T1 tumor-bearing mouse model of large volume. (B) Distant tumor growth curves of mice after different treatments ( n = 5). (C) In vitro images of the distant tumors ( n = 5). (D) In vitro distant tumors mass ( n = 5). (E) Immunohistochemical staining of CD3 in tumor tissue slices after different treatments (scale bars = 100 μm). The images below were the corresponding enlarged parts (scale bars = 25 μm). (F) Immunohistochemical staining of CD8 in tumor tissue slices after various treatments (scale bars = 100 μm). The images below were the corresponding enlarged parts (scale bars = 25 μm). Data are presented as mean ± SD. ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 determined by Student’s t -test.

Article Snippet: InVivo MAb anti-mouse PD-L1 was purchased from Bioxcell (USA).

Techniques: In Vitro, Immunohistochemical staining, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Activation of Carbonic Anhydrase IX by Alternatively Spliced Tissue Factor Under Late-Stage Tumor Conditions

doi: 10.1038/labinvest.2016.103

Figure Lengend Snippet: A) Cell culture conditions for modeling early stage or late stage pancreatic cancer progression. B) Expression of the hypoxia-associated proteins HIF-1α (117 kD), HIF-2α (118 kD), and CAIX (55 kD) in early (norm) or late stage (hyp) environments. β-Actin served as a loading control. HeLa cells treated with 100 μM CoCl 2 were used as a positive control for hypoxia-induced gene expression. “env,” the environment under which the cells were kept for 48 hours before lysing, with “hyp” indicating hypoxic/low glucose and “norm” indicating normoxic/high glucose. The bottom panel shows the over-expression of asTF in Pt45.P1 cells, vimentin , and the loading control β-Actin.

Article Snippet: The separated proteins were then transferred to a PVDF membrane, blocked with 5% nonfat milk, probed with antibodies to asTF (rabbit monoclonal RabMab1) , vimentin (rabbit mAb, Cell Signaling), β-actin (rabbit, Cell Signaling), HIF-1α (rabbit pAb, Bethyl Laboratories), HIF-2α (rabbit pAb, GeneTex) CAIX (mouse mAb, GeneTex), or CAXII (goat pAb, Acris), followed by probing with the corresponding HRP-conjugated secondary antibodies (Bio-Rad) and developed using ECL and H 2 O 2 .

Techniques: Cell Culture, Expressing, Control, Positive Control, Gene Expression, Over Expression

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: 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