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Proteintech cbx3 polyclonal antibody
Significant changes of CBX expression in the transcription level between ESCA and normal esophagus tissues (ONCOMINE).

Significant changes of CBX expression in the transcription level between ESCA and normal esophagus tissues (ONCOMINE).

Cbx3 Polyclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cbx3+polyclonal+antibody/CBX3+Antibody/pmc09019303-82-15-18
Average 93 stars, based on 30 article reviews

cbx3 polyclonal antibody - by Bioz Stars, 2026-09

93/100 stars

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1) Product Images from "Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer"

Article Title: Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer

Journal: Frontiers in Genetics

doi: 10.3389/fgene.2022.851390

Figure Legend Snippet: Significant changes of CBX expression in the transcription level between ESCA and normal esophagus tissues (ONCOMINE).

Techniques Used: Expressing

Protein expression levels of CBX1, CBX3, CBX7, and CBX8 in 12 ESCA tissues and adjacent normal tissues (western blot). (A) CBX1, CBX3, and CBX8 were upregulated in ESCA tissues compared with adjacent normal tissues, while CBX7 was expressed at similar levels in ESCA and normal esophageal tissues. (B–E) Statistical results of western blotting of CBX1, CBX3, CBX7, and CBX8 in ESCA tissues and normal tissues. (* p < 0.05, analysis by paired t-test.)

Figure Legend Snippet: Protein expression levels of CBX1, CBX3, CBX7, and CBX8 in 12 ESCA tissues and adjacent normal tissues (western blot). (A) CBX1, CBX3, and CBX8 were upregulated in ESCA tissues compared with adjacent normal tissues, while CBX7 was expressed at similar levels in ESCA and normal esophageal tissues. (B–E) Statistical results of western blotting of CBX1, CBX3, CBX7, and CBX8 in ESCA tissues and normal tissues. (* p < 0.05, analysis by paired t-test.)

Techniques Used: Expressing, Western Blot

Fluorescence images of paraffin-embedded sections of human ESCA and adjacent tissues with confocal microscopy. (A) CBX1 was significantly higher in ESCA tissues than that in precancerous tissues. (B) CBX3 was significantly higher in ESCA tissues than that in precancerous tissues. (C) CBX8 was significantly higher in ESCA tissues than that in precancerous tissues. The white dashed lines separate the precancerous tissues (on the left side of the figure) from the ESCA tissues (on the right side of the figure). Scale bars = 25 um. (Magnification 63✕/1.40 oil).

Figure Legend Snippet: Fluorescence images of paraffin-embedded sections of human ESCA and adjacent tissues with confocal microscopy. (A) CBX1 was significantly higher in ESCA tissues than that in precancerous tissues. (B) CBX3 was significantly higher in ESCA tissues than that in precancerous tissues. (C) CBX8 was significantly higher in ESCA tissues than that in precancerous tissues. The white dashed lines separate the precancerous tissues (on the left side of the figure) from the ESCA tissues (on the right side of the figure). Scale bars = 25 um. (Magnification 63✕/1.40 oil).

Techniques Used: Fluorescence, Confocal Microscopy

Expressing:

Article Title: Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer
Article Snippet: Proteins were separated by SDS-PAGE and were visualized using the ECL system (Bio-Rad, Hercules, CA, United States).Proteins were separated by SDS-PAGE and were visualized using the ECL system (Bio-Rad, Hercules, CA, United States).. Primary antibodies used in this study were as follows: CBX1 Polyclonal Antibody (Proteintech, 10241-2 -AP), CBX3 Polyclonal Antibody (Proteintech, 11650-2 -AP), CBX7 Polyclonal Antibody (Proteintech, 26278-1-AP), and CBX8 Monoclonal Antibody (Santa Cruz, sc-374332).. Monoclonal Anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States).Monoclonal Anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States).

Western Blot:

Fluorescence:

Confocal Microscopy:

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Figure 1. Identiﬁcation of MC antigens reactive to IgA auto-Abs generated in gddY mice. (A) Detection of antigens for rmAbs derived from gddY mouse PBs in the lysates of gddY mouse primary MCs by WB. Split membranes, each containing proteins in one lane, were incubated with the indicated rmAbs (1 μg/ml), followed by an anti-human IgG Ab. (B) Immunoprecipitation of gddY mouse MC lysates with rmAb#66 or the control rmAb#NP. SDS–PAGE of the precipitates was developed using silver staining. The bands indicated by arrows were excised and analyzed using mass spectrometry. Those from the rmAb#66 precipitate included CBX1, CBX3, and CBX5, as indicated by arrows. (C) FLAG-tagged CBX1, CBX3, or CBX5 proteins transiently expressed in HEK293T cells were detected by WB with the indicated rmAbs (1 μg/ml), followed by anti-human IgG Ab (left panel) or anti-FLAG Ab (right panel). (D) ELISA screening of rmAbs for binding to CBX proteins. The heatmap represents the resulting OD values of the indicated rmAbs, with “+” indicating a saturated signal. (E) Reactivity of gddY mouse serum IgA with the CBX proteins. WB was performed as in (C) using sera of individual BALB/c or gddY mice as the primary Ab as indicated and anti-IgA Ab as the secondary Ab (left, top) or anti-FLAG Ab to conﬁrm the presence of CBX proteins on the blot (left, bottom). Representative data are shown here. Venn diagram showing the percentage of gddY mice whose serum IgA reacted with CBX1, CBX3, or CBX5 (right panel) (n = 14). (F) ELISA determination of anti-CBX3 IgA Abs in sera from healthy controls (HC; n = 30) and patients with IgAN (IgAN; n = 70). ***: 0.0006 (Mann–Whitney U test). The dashed line shows 99% conﬁdence interval (CI) of the control sera. One of two or three independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Journal: Life science alliance

Article Title: Oral bacteria induce IgA autoantibodies against a mesangial protein in IgA nephropathy model mice.

doi: 10.26508/lsa.202402588

Figure Lengend Snippet: Figure 1. Identiﬁcation of MC antigens reactive to IgA auto-Abs generated in gddY mice. (A) Detection of antigens for rmAbs derived from gddY mouse PBs in the lysates of gddY mouse primary MCs by WB. Split membranes, each containing proteins in one lane, were incubated with the indicated rmAbs (1 μg/ml), followed by an anti-human IgG Ab. (B) Immunoprecipitation of gddY mouse MC lysates with rmAb#66 or the control rmAb#NP. SDS–PAGE of the precipitates was developed using silver staining. The bands indicated by arrows were excised and analyzed using mass spectrometry. Those from the rmAb#66 precipitate included CBX1, CBX3, and CBX5, as indicated by arrows. (C) FLAG-tagged CBX1, CBX3, or CBX5 proteins transiently expressed in HEK293T cells were detected by WB with the indicated rmAbs (1 μg/ml), followed by anti-human IgG Ab (left panel) or anti-FLAG Ab (right panel). (D) ELISA screening of rmAbs for binding to CBX proteins. The heatmap represents the resulting OD values of the indicated rmAbs, with “+” indicating a saturated signal. (E) Reactivity of gddY mouse serum IgA with the CBX proteins. WB was performed as in (C) using sera of individual BALB/c or gddY mice as the primary Ab as indicated and anti-IgA Ab as the secondary Ab (left, top) or anti-FLAG Ab to conﬁrm the presence of CBX proteins on the blot (left, bottom). Representative data are shown here. Venn diagram showing the percentage of gddY mice whose serum IgA reacted with CBX1, CBX3, or CBX5 (right panel) (n = 14). (F) ELISA determination of anti-CBX3 IgA Abs in sera from healthy controls (HC; n = 30) and patients with IgAN (IgAN; n = 70). ***: 0.0006 (Mann–Whitney U test). The dashed line shows 99% conﬁdence interval (CI) of the control sera. One of two or three independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Article Snippet: For FCM, the following Abs were used: anti-mouse IgA-APC (clone 11- 44-2; Southern Biotech), anti-mouse CD73-biotin (clone TY/11.8; BioLegend), anti-mouse CD45-PerCP-Cy5.5 (clone 30-F11; BioLegend), anti-mouse CD31-PE (clone 390; BioLegend), anti-mouse TER119BV510 (clone TER-119; BioLegend), rabbit polyclonal anti-HP1γ (CBX3) Ab (2619; Cell Signaling Technology), rabbit monoclonal anti-HP1β (CBX1) Ab (8676; Cell Signaling Technology), rabbit IgG (for isotypematched control; 011-000-003; Jackson ImmunoResearch), antirabbit IgG-BV421 (clone Poly4064; BioLegend), and anti-human IgG Fcγ-Alexa Fluor 647 (709-606-098; Jackson ImmunoResearch).

Techniques: Generated, Derivative Assay, Incubation, Immunoprecipitation, Control, SDS Page, Silver Staining, Mass Spectrometry, Enzyme-linked Immunosorbent Assay, Binding Assay, MANN-WHITNEY

Figure 2. CBX3 is an autoantigen expressed on the surfaces of MCs. (A, B) Representative histograms of FCM analysis of glomerular CD45– cells from BALB/c mice stained with Abs against CD45, CD73, and CD31 for gating on MCs (CD73+

Journal: Life science alliance

Article Title: Oral bacteria induce IgA autoantibodies against a mesangial protein in IgA nephropathy model mice.

doi: 10.26508/lsa.202402588

Figure Lengend Snippet: Figure 2. CBX3 is an autoantigen expressed on the surfaces of MCs. (A, B) Representative histograms of FCM analysis of glomerular CD45– cells from BALB/c mice stained with Abs against CD45, CD73, and CD31 for gating on MCs (CD73+

Techniques: Staining

Figure 5. The gddY auto-Abs reacted with commensal bacteria in the oral cavity of the gddY mice. (A, B) WB analysis of lysates of bacteria from feces (fecal), small intestine (SI), and oral cavity (oral) of gddY mice blotted with sera from gddY mice (A) or culture supernatant of leukocytes from gddY mouse kidneys (B) as the primary Abs and anti-IgA as the secondary Ab. The IgA concentration in the primary Abs was adjusted to 1 μg/ml by dilution. The bacteria were cultured anaerobically on brain heart infusion agar before lysis. (C, D, E, F) FCM analyses of cultured bacteria stained with various Ab solutions (containing 1 μg/ml IgA) and SYBR Green (for bacterial DNA). (C, D) Bacteria from the oral cavity (oral) or small intestine (C), or oral bacteria from BALB/c or gddY mice (D), were stained with sera from gddY mice. (E) Oral bacteria from gddY mice were stained with the culture supernatant of leukocytes from the indicated organs of gddY mice. (C, D, E) APC-conjugated anti-mouse IgA was used as secondary Ab. The secondary Ab alone did not stain cultured bacteria (data not shown). (F) Oral or fecal bacteria from gddY mice cultured under the indicated conditions were stained with rmAb#66 or rmAb#NP, followed by Alexa Fluor 647–anti-human IgG. (C, D, E) The frequencies of IgA-bound bacteria among whole bacteria stained with SYBR Green are plotted. Data are presented as means ± SD of biological replicates. (G) rmAb#66 (1 μg/ml) was incubated with the indicated concentrations of recombinant CBX3 or OVA in equal volumes for 20 min at RT, mixed with cultured oral bacteria from gddY mice, and incubated overnight on ice. The bacteria were stained with Alexa Fluor 647-anti-human IgG to detect bound rmAb#66, together with SYBR Green to label the bacteria, and analyzed by FCM. The frequencies of rmAb#66-bound bacteria among all bacteria were plotted. Data are presented as means ± SD of three technical replicates. *P < 0.05; ***P < 0.005; ****P < 0.001. P-values were calculated using unpaired t test. One of two independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Journal: Life science alliance

Article Title: Oral bacteria induce IgA autoantibodies against a mesangial protein in IgA nephropathy model mice.

doi: 10.26508/lsa.202402588

Figure Lengend Snippet: Figure 5. The gddY auto-Abs reacted with commensal bacteria in the oral cavity of the gddY mice. (A, B) WB analysis of lysates of bacteria from feces (fecal), small intestine (SI), and oral cavity (oral) of gddY mice blotted with sera from gddY mice (A) or culture supernatant of leukocytes from gddY mouse kidneys (B) as the primary Abs and anti-IgA as the secondary Ab. The IgA concentration in the primary Abs was adjusted to 1 μg/ml by dilution. The bacteria were cultured anaerobically on brain heart infusion agar before lysis. (C, D, E, F) FCM analyses of cultured bacteria stained with various Ab solutions (containing 1 μg/ml IgA) and SYBR Green (for bacterial DNA). (C, D) Bacteria from the oral cavity (oral) or small intestine (C), or oral bacteria from BALB/c or gddY mice (D), were stained with sera from gddY mice. (E) Oral bacteria from gddY mice were stained with the culture supernatant of leukocytes from the indicated organs of gddY mice. (C, D, E) APC-conjugated anti-mouse IgA was used as secondary Ab. The secondary Ab alone did not stain cultured bacteria (data not shown). (F) Oral or fecal bacteria from gddY mice cultured under the indicated conditions were stained with rmAb#66 or rmAb#NP, followed by Alexa Fluor 647–anti-human IgG. (C, D, E) The frequencies of IgA-bound bacteria among whole bacteria stained with SYBR Green are plotted. Data are presented as means ± SD of biological replicates. (G) rmAb#66 (1 μg/ml) was incubated with the indicated concentrations of recombinant CBX3 or OVA in equal volumes for 20 min at RT, mixed with cultured oral bacteria from gddY mice, and incubated overnight on ice. The bacteria were stained with Alexa Fluor 647-anti-human IgG to detect bound rmAb#66, together with SYBR Green to label the bacteria, and analyzed by FCM. The frequencies of rmAb#66-bound bacteria among all bacteria were plotted. Data are presented as means ± SD of three technical replicates. *P < 0.05; ***P < 0.005; ****P < 0.001. P-values were calculated using unpaired t test. One of two independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Techniques: Bacteria, Concentration Assay, Cell Culture, Lysis, Staining, SYBR Green Assay, Incubation, Recombinant

Figure 6. Identiﬁcation of a bacterial strain that shares an rmAb#66 epitope with CBX3. (A) Isolated bacteria (C42) were stained with rmAb#66 or rmAb#NP (1 μg/ml), followed by Alexa Fluor 647–anti-human IgG together with SYBR Green, and analyzed by FCM. (B) rmAb#66 (1 μg/ml) was incubated with CBX3 or OVA and then with C42, which was stained with anti-human IgG and analyzed as described in Fig 5G. Data are presented as means ± SD of two technical replicates. (C) C42 bacteria (2 × 106 cells) were incubated with the indicated concentrations of PNGase F (Roche) for 4 h at 37°C, stained with rmAb#66 (1 μg/ml) overnight on ice, and labeled with SYBR Green. Binding of rmAb#66 to C42 was detected by FCM using Alexa Fluor 647-anti-human IgG (n = 2). (D) BALB/c mice were immunized s.c. with UV-killed C42 or E. faecalis, together with zymosan as an adjuvant, at 4 wk of age. WB analysis was performed as shown in Fig 3E (except that CBX1 and CBX5 samples were omitted) with sera from immunized mice at 8 wk of age as the primary Abs and anti-IgG Ab as the secondary Ab (left panel) or anti-FLAG Ab (right panel). (E) Representative IFM images of kidney sections from the mice used in (D). Kidney sections were stained with an anti-IgA Ab (red). Dashed circles indicate areas of glomeruli and white lines indicate scale bars (100 μm). (F) Frequency of bacteria recognized by rmAb#66 among all bacteria (SYBR Green+) in the oral cavity of gddY mice (n = 6). Bacteria from gddY mice fed a normal diet (control) or an ED (ED) were cultured anaerobically on brain heart infusion agar, stained with rmAb#66 (1 μg/ml) and Alexa Fluor 647–anti-human IgG, together with SYBR Green, and analyzed by FCM. (G) Frequency of each bacterial strain among the total bacteria in the oral cavities of gddY mice fed a normal diet (control) or an ED, as determined by 16S rRNA bulk sequencing (n = 6). An underscore after each genus name indicates that the species was not classiﬁed. Data are presented as means ± SD of biological replicates. *P < 0.05; ***P < 0.005; ****P < 0.001. P-values were calculated using the unpaired t test. One of two independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Journal: Life science alliance

Article Title: Oral bacteria induce IgA autoantibodies against a mesangial protein in IgA nephropathy model mice.

doi: 10.26508/lsa.202402588

Figure Lengend Snippet: Figure 6. Identiﬁcation of a bacterial strain that shares an rmAb#66 epitope with CBX3. (A) Isolated bacteria (C42) were stained with rmAb#66 or rmAb#NP (1 μg/ml), followed by Alexa Fluor 647–anti-human IgG together with SYBR Green, and analyzed by FCM. (B) rmAb#66 (1 μg/ml) was incubated with CBX3 or OVA and then with C42, which was stained with anti-human IgG and analyzed as described in Fig 5G. Data are presented as means ± SD of two technical replicates. (C) C42 bacteria (2 × 106 cells) were incubated with the indicated concentrations of PNGase F (Roche) for 4 h at 37°C, stained with rmAb#66 (1 μg/ml) overnight on ice, and labeled with SYBR Green. Binding of rmAb#66 to C42 was detected by FCM using Alexa Fluor 647-anti-human IgG (n = 2). (D) BALB/c mice were immunized s.c. with UV-killed C42 or E. faecalis, together with zymosan as an adjuvant, at 4 wk of age. WB analysis was performed as shown in Fig 3E (except that CBX1 and CBX5 samples were omitted) with sera from immunized mice at 8 wk of age as the primary Abs and anti-IgG Ab as the secondary Ab (left panel) or anti-FLAG Ab (right panel). (E) Representative IFM images of kidney sections from the mice used in (D). Kidney sections were stained with an anti-IgA Ab (red). Dashed circles indicate areas of glomeruli and white lines indicate scale bars (100 μm). (F) Frequency of bacteria recognized by rmAb#66 among all bacteria (SYBR Green+) in the oral cavity of gddY mice (n = 6). Bacteria from gddY mice fed a normal diet (control) or an ED (ED) were cultured anaerobically on brain heart infusion agar, stained with rmAb#66 (1 μg/ml) and Alexa Fluor 647–anti-human IgG, together with SYBR Green, and analyzed by FCM. (G) Frequency of each bacterial strain among the total bacteria in the oral cavities of gddY mice fed a normal diet (control) or an ED, as determined by 16S rRNA bulk sequencing (n = 6). An underscore after each genus name indicates that the species was not classiﬁed. Data are presented as means ± SD of biological replicates. *P < 0.05; ***P < 0.005; ****P < 0.001. P-values were calculated using the unpaired t test. One of two independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Techniques: Isolation, Bacteria, Staining, SYBR Green Assay, Incubation, Labeling, Binding Assay, Adjuvant, Control, Cell Culture, Sequencing

Figure 7. Determination of rmAb#66 epitope on CBX3. (A) Schematic representation of CBX3 proteins: 1/2 CBX3 (AA1-92) and 2/3 CBX3 (AA63-184). (B, C) FLAG-tagged 1/2 or 2/3 CBX3 proteins, transiently expressed in HEK293T cells, were detected by WB with rmAb#66 or rmAb#NP (1 μg/ml) followed by anti-human IgG Ab ((B), left panel). The same membrane was reblotted with sera of two gddY mice followed by anti-mouse IgA Ab ((C), left panel). Protein loading was conﬁrmed using an anti-FLAG Ab ((B, C), right panels; the same blot is shown). (D) Amino acid sequence alignment of CBX proteins (AA1-62) (top). WB analysis of FLAG-tagged WT or the indicated mutated CBX3 proteins transiently expressed in HEK293T cells, performed as in (B) (bottom). (E) Reactivity to CBX3 (WT) or CBX3 mutant 54FA (54FA) proteins of the indicated rmAbs at the indicated concentrations was evaluated by ELISA and presented as OD450 values. One of two independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Journal: Life science alliance

Article Title: Oral bacteria induce IgA autoantibodies against a mesangial protein in IgA nephropathy model mice.

doi: 10.26508/lsa.202402588

Figure Lengend Snippet: Figure 7. Determination of rmAb#66 epitope on CBX3. (A) Schematic representation of CBX3 proteins: 1/2 CBX3 (AA1-92) and 2/3 CBX3 (AA63-184). (B, C) FLAG-tagged 1/2 or 2/3 CBX3 proteins, transiently expressed in HEK293T cells, were detected by WB with rmAb#66 or rmAb#NP (1 μg/ml) followed by anti-human IgG Ab ((B), left panel). The same membrane was reblotted with sera of two gddY mice followed by anti-mouse IgA Ab ((C), left panel). Protein loading was conﬁrmed using an anti-FLAG Ab ((B, C), right panels; the same blot is shown). (D) Amino acid sequence alignment of CBX proteins (AA1-62) (top). WB analysis of FLAG-tagged WT or the indicated mutated CBX3 proteins transiently expressed in HEK293T cells, performed as in (B) (bottom). (E) Reactivity to CBX3 (WT) or CBX3 mutant 54FA (54FA) proteins of the indicated rmAbs at the indicated concentrations was evaluated by ELISA and presented as OD450 values. One of two independent experiments with similar results is shown in each panel. Source data are available for this ﬁgure.

Techniques: Membrane, Sequencing, Mutagenesis, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Genetics

Article Title: Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer

doi: 10.3389/fgene.2022.851390

Figure Lengend Snippet: Significant changes of CBX expression in the transcription level between ESCA and normal esophagus tissues (ONCOMINE).

Article Snippet: Primary antibodies used in this study were as follows: CBX1 Polyclonal Antibody (Proteintech, 10241-2 -AP), CBX3 Polyclonal Antibody (Proteintech, 11650-2 -AP), CBX7 Polyclonal Antibody (Proteintech, 26278-1-AP), and CBX8 Monoclonal Antibody (Santa Cruz, sc-374332).

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer

doi: 10.3389/fgene.2022.851390

Figure Lengend Snippet: Protein expression levels of CBX1, CBX3, CBX7, and CBX8 in 12 ESCA tissues and adjacent normal tissues (western blot). (A) CBX1, CBX3, and CBX8 were upregulated in ESCA tissues compared with adjacent normal tissues, while CBX7 was expressed at similar levels in ESCA and normal esophageal tissues. (B–E) Statistical results of western blotting of CBX1, CBX3, CBX7, and CBX8 in ESCA tissues and normal tissues. (* p < 0.05, analysis by paired t-test.)

Techniques: Expressing, Western Blot

Journal: Frontiers in Genetics

Article Title: Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer

doi: 10.3389/fgene.2022.851390

Figure Lengend Snippet: Fluorescence images of paraffin-embedded sections of human ESCA and adjacent tissues with confocal microscopy. (A) CBX1 was significantly higher in ESCA tissues than that in precancerous tissues. (B) CBX3 was significantly higher in ESCA tissues than that in precancerous tissues. (C) CBX8 was significantly higher in ESCA tissues than that in precancerous tissues. The white dashed lines separate the precancerous tissues (on the left side of the figure) from the ESCA tissues (on the right side of the figure). Scale bars = 25 um. (Magnification 63✕/1.40 oil).

Techniques: Fluorescence, Confocal Microscopy

cDNAs of the differentially expressed genes were identified and non-quantitative RT-PCR was used to examine their expression levels in gonads. Homologs of mouse and zebrafish were also analyzed for comparision. Myosin, Ferritin, Cbx3 are highly expressed in testis; Glutathione S-transferase M, triosephosphate isomerase B are highly expressed in ovotestis. Rab37 and Pentraxin are highly expressed in ovary. Protein expression levels of three spots for each of these genes are listed in Fig. S4. Some of the genes have no difference between gonads, like Rab1A and Enolase. PCR product for each gene has been run under the same experimental conditions and same sizes of gel images have been used for all genes.

Journal: Scientific Reports

Article Title: Proteomic analysis of three gonad types of swamp eel reveals genes differentially expressed during sex reversal

doi: 10.1038/srep10176

Figure Lengend Snippet: cDNAs of the differentially expressed genes were identified and non-quantitative RT-PCR was used to examine their expression levels in gonads. Homologs of mouse and zebrafish were also analyzed for comparision. Myosin, Ferritin, Cbx3 are highly expressed in testis; Glutathione S-transferase M, triosephosphate isomerase B are highly expressed in ovotestis. Rab37 and Pentraxin are highly expressed in ovary. Protein expression levels of three spots for each of these genes are listed in Fig. S4. Some of the genes have no difference between gonads, like Rab1A and Enolase. PCR product for each gene has been run under the same experimental conditions and same sizes of gel images have been used for all genes.

Article Snippet: Anti-β-actin and anti-Cbx3 polyclonal antibodies were purchased from Santa Cruz Biotech (CA, USA).

Techniques: Quantitative RT-PCR, Expressing

a ). Protein sequence alignments of Cbx3 from human, mouse, cattle, chicken, frog, salmon, zebrafish and swamp eel show an evolutionarily conservation. GenBank access number for swamp eel Cbx3 is KP054389. b ) Phylogenetic tree depicts the Cbx family from human, mouse, cattle, chicken, frog, salmon, zebrafish and swamp eel. The numbers on the branches represent the boot-strap values from 100 replicates obtained using the maximum-likelihood method. The red color highlighs the Cbx3 cluster. c ) Non-quantitative RT-PCR analysis in adult tissues from each sex of swamp eel. PCR product for each gene has been run under the same experimental conditions and same sizes of gel images have been used for all genes. d ) Western blot assay shows Cbx3 expression in three gonad types of swamp eel. The protein gels have been run under the same experimental conditions and same sizes of gel images have been used for all proteins. The right panel shows the intensity values related to Actin. e ) Immunohistochemistry assay shows that Cbx3 expression in gonads of swamp eel. Marked signals are observed in testis and then in ovotestis. The signals are hardly detected in ovary. O, ovary, Ode, developing ovary. f ) Immunofluorescence analysis of Cbx3 in mouse testis using anti-Cbx3. Signals are observed in spermatogonia (sg), spermatocytes (sc) and sperm cells (s). The bar, 20 μm.

Journal: Scientific Reports

Article Title: Proteomic analysis of three gonad types of swamp eel reveals genes differentially expressed during sex reversal

doi: 10.1038/srep10176

Figure Lengend Snippet: a ). Protein sequence alignments of Cbx3 from human, mouse, cattle, chicken, frog, salmon, zebrafish and swamp eel show an evolutionarily conservation. GenBank access number for swamp eel Cbx3 is KP054389. b ) Phylogenetic tree depicts the Cbx family from human, mouse, cattle, chicken, frog, salmon, zebrafish and swamp eel. The numbers on the branches represent the boot-strap values from 100 replicates obtained using the maximum-likelihood method. The red color highlighs the Cbx3 cluster. c ) Non-quantitative RT-PCR analysis in adult tissues from each sex of swamp eel. PCR product for each gene has been run under the same experimental conditions and same sizes of gel images have been used for all genes. d ) Western blot assay shows Cbx3 expression in three gonad types of swamp eel. The protein gels have been run under the same experimental conditions and same sizes of gel images have been used for all proteins. The right panel shows the intensity values related to Actin. e ) Immunohistochemistry assay shows that Cbx3 expression in gonads of swamp eel. Marked signals are observed in testis and then in ovotestis. The signals are hardly detected in ovary. O, ovary, Ode, developing ovary. f ) Immunofluorescence analysis of Cbx3 in mouse testis using anti-Cbx3. Signals are observed in spermatogonia (sg), spermatocytes (sc) and sperm cells (s). The bar, 20 μm.

Article Snippet: Anti-β-actin and anti-Cbx3 polyclonal antibodies were purchased from Santa Cruz Biotech (CA, USA).

Techniques: Sequencing, Quantitative RT-PCR, Western Blot, Expressing, Immunohistochemistry, Immunofluorescence

The color of nodes represents expression level relative to Actin among ovary, ovotestis and testis as the color panel indicated. Protein relationships are obtained from BioGrid. All proteins are the swamp eel homologs. Rab37, Cbx3 and Pax3 are identified in the MS analysis.

Journal: Scientific Reports

Article Title: Proteomic analysis of three gonad types of swamp eel reveals genes differentially expressed during sex reversal

doi: 10.1038/srep10176

Figure Lengend Snippet: The color of nodes represents expression level relative to Actin among ovary, ovotestis and testis as the color panel indicated. Protein relationships are obtained from BioGrid. All proteins are the swamp eel homologs. Rab37, Cbx3 and Pax3 are identified in the MS analysis.

Article Snippet: Anti-β-actin and anti-Cbx3 polyclonal antibodies were purchased from Santa Cruz Biotech (CA, USA).

Techniques: Expressing

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cbx3 polyclonal antibody (Proteintech)

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1) Product Images from "Identification and Validation of Chromobox Family Members as Potential Prognostic Biomarkers and Therapeutic Targets for Human Esophageal Cancer"

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