FITC anti-mouse CD5 100605 from BioLegend

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Development (Cambridge, England)
January 31 2018

Fig 1: Attenuation of Hh signalling by rHhip treatment of WT and Gli3-/- FTOCs. (A-G) WT (n=4) and Gli3-/- (n=6) E17.5 FTOCs were treated with rHhip or left untreated (control) for 4 days, and analysed by flow cytometry. (B,D,F,G) Scatter plots show relative mean±s.e.m., where each point represents the percentage of SP4 (circles), SP8 (squares) and DP (crosses) populations of one rHhip-treated lobe from an individual thymus, divided by the percentage of that population from the untreated lobe from the same thymus. (A,C) Dot plots of (A) CD8 versus CD4 and (C) CD8 versus CD4 gated on CD3hi. (B,D) Percentage of rHhip-treated populations, giving significance by Student's t-test compared with untreated control for (B) SP4 (WT, P<0.05; Gli3-/-, P<0.03), SP8 (WT, P<0.002), DP (WT, P<0.006); and for (D) CD3hi SP4 (WT, P<0.04; Gli3-/-, P<0.05), CD3hi SP8 (WT, P<0.05), CD3hi DP (WT, P<0.05). (E) Scatter plots of (left) MFI of CD5 on SP4, DP and SP8 cells and (right) in the same populations gated on CD3hi from a representative experiment. (F) MFI of CD5 in Hhip-treated FTOC on SP4, SP8 and DP showing significance by Student's t-test compared with untreated control for SP4 (WT, P<0.05); and in (G) MFI of CD5 showing significance by Student's t-test compared with untreated control on CD3hi SP4 (WT, P<0.002), CD3hi SP8 (WT, P<0.0001) and CD3hi DP (Gli3-/-, P<0.05).
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Development (Cambridge, England)
January 31 2018

Fig 2: Gli3 expression in TECs and not thymocyte-intrinsic Gli3 expression is required for SP4 development. (A-D) Flow cytometry of fresh E17.5 Gli3fl/fl VavCre- control (n=7) and Gli3fl/fl VavCre+ (n=6) thymus. Scatter plots show relative mean±s.e.m., where each point represents thymus from a different embryo for control littermate and Gli3fl/fl VavCre+. (A) CD4 and CD8 expression. (B) Relative percentage of thymocyte subsets. (C) Relative MFI of anti-CD5 staining on thymocyte subsets. (D) Representative histogram showing anti-CD5 staining on the DP population in control (solid line) and Gli3fl/fl VavCre+ (dotted line), showing MFIs. No significant differences were found between control and Cre+ thymi. (E-H) Flow cytometry of fresh E17.5 thymus from Gli3fl/fl FoxN1Cre- (control, n=4) and Gli3fl/fl FoxN1Cre+ (n=4) littermates. Scatter plots show relative mean±s.e.m., where each point represents thymus from a different embryo for control and Gli3fl/fl FoxN1Cre+. (E) CD8 against CD4. (F) Relative mean percentage of SP4, DP, SP8 and DN populations, giving significance by Student's t-test compared with control littermate for SP4 (P<0.01), DP (P<0.05) and DN (P<0.04). (G) Representative histogram shows CD5 staining on DP thymocytes from control (Cre-, solid line) and Cre+ (dotted line). MFI of CD5 fluorescence is given. (H) Relative mean MFI of CD5 on SP4, DP, SP8 and DN populations. Differences are significant between Cre+ and control for DP (P<0.03) and DN (P<0.05). (I-M) Flow cytometry analysis of E17.5 FTOC +4 days from Gli3fl/fl FoxN1Cre- (n=8) and Gli3fl/fl FoxN1Cre+ (n=10) littermates. Scatter plots show relative mean±s.e.m., where each point represents thymus from a different embryo for control FTOC and Gli3fl/fl FoxN1Cre+ FTOC. (I) CD8 against CD4. (J) CD8 against CD4, gated on CD3hi. (K) Relative percentage of SP4, DP and SP8 populations and SP4:SP8 ratio, with significance by Student's t-test compared with Cre- littermate for SP4 (P<0.02), DP (P<0.007) and SP4:SP8 ratio (P<0.05). (L) Relative percentage of thymocyte populations and CD4:CD8 ratio, gated on CD3hi, with significance compared with Cre- littermate, for CD3hi SP4 (P<0.05) and CD3hi SP4:SP8 ratio (P<0.04). (M) Relative MFI of CD5 on thymocyte populations, with significance compared with Cre- for DP (P<0.007). (N,O) Flow cytometry of E17.5 FTOC +4 days from control Gli3fl/fl VavCre- (n=7) and Gli3fl/fl VavCre+ (n=6) littermates. No significant differences were found between Cre- and Cre+. (N) CD4 against CD8. (O) CD4 against CD8, gated on CD3hi. (P) Relative percentage of SP4, DP and SP8 populations and SP4:SP8 ratio. (Q) Gated on CD3hi, relative percentage of SP4, DP and SP8 populations and SP4:SP8 ratio. (R) MFI of CD5, gated on thymocyte populations.
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Development (Cambridge, England)
January 31 2018

Fig 3: Microarray and QRT-PCR analyses of transcriptional differences in thymic selection genes in sorted populations from WT and Gli3 mutant thymus and in rShh-treated WT FTOC. (A,B) Normalised microarray transcript expression in Gli3-/- and WT datasets of (A) Egr2 and (B) Tox. (C,D) QRT-PCR analysis showing a representative experiment (of three) for expression of (C) Egr2 and (D) Tox in thymocytes prepared from E17.5 WT control and rShh-treated FTOCs for 2 days. Differences between control and rShh treated were significant: Egr2 (P<0.02), Tox (P<0.07). Transcripts were normalised relative to Hprt. (E) Normalised microarray transcript expression in Gli3-/- and WT datasets of Nr4a1 (top) and Cd5 (bottom). (F) CCA showing the separation of the WT, Gli3+/- (HET) and Gli3-/- (KO) SP4 microarray datasets on a scale of strong to weak TCR stimulation.
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Development (Cambridge, England)
January 31 2018

Fig 4: Neutralisation of Hh in Gli3fl/flFoxN1Cre+ FTOC and conditional deletion of Shh from TECs increase SP4 differentiation. (A-L) Flow cytometry analysis with scatter plots showing relative mean±s.e.m., where each point represents thymus from a different embryo. (A-E) E17.5 Gli3fl/fl FoxN1Cre+ FTOC treated with rHhip for 4 days (n=7) compared with the untreated lobe (control) from the same thymus (n=7). (A) CD8 against CD4. (B) Percentage of thymocyte populations, showing significance by Student's t-test for rHhip treated versus control untreated for SP4 (P<0.02) and DN (P<0.006). (C) The percentage of DP thymocytes, gated on CD3hi. The difference in percentage of CD3hi DP cells between rHhip treated and control was statistically significant by Student's t-test (P<0.02). (D) CD8 versus CD4, gated on CD3hi. (E) MFI of CD5 on rHhip-treated FTOC showing significance by Student's t-test versus control untreated FTOC for SP4 (P<0.02) and DP (P<0.007). (F) Gated on CD3hi, MFI of CD5 on rHhip treated showing significance by Student's t-test versus control untreated FTOC for CD3hi SP4 (P<0.01) and CD3hi DP (P<0.02). (G-I) Flow cytometry of E17.5 FTOC +4 days from Shhfl/fl FoxN1Cre- (n=4, control) and Shhfl/fl FoxN1Cre+ (n=6) littermates. Scatter plots show relative mean±s.e.m., where each point represents thymus from a different embryo. (H) Percentage of thymocyte populations and SP4:SP8 ratio, in control and Shhfl/fl FoxN1Cre+, giving significance by Student's t-test compared with Cre- for SP4 (P<0.02), DP (P<0.04) and SP4:SP8 ratio (P<0.05). (I) Percentage of thymocyte populations and SP4:SP8 ratio gated on CD3hi, in control and Shhfl/fl FoxN1Cre+, giving significance by Student's t-test compared with Cre- for CD3hi SP4 (P<0.003) and CD3hi DP (P<0.002). (J-L) E17.5 Shhfl/fl FoxN1Cre+ FTOC treated with 1 µg/ml rShh for 4 days compared with control untreated Shhfl/fl FoxN1Cre+ FTOC thymus lobe from the same thymus (n=5). (J) CD8 against CD4 staining (top); CD8 against CD4 staining, gated on CD3hi (bottom). (K) Percentage of thymocyte population, divided by the percentage from the control untreated cultured lobe from the same thymus, giving significance compared with untreated for SP4 (P<0.04). (L) Percentage of thymocyte population, gated on CD3hi, divided by the percentage from the control untreated cultured lobe from the same thymus, giving significance compared with untreated control for CD3hi DP (P<0.02) and CD3hi SP8 (P<0.03).
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Development (Cambridge, England)
January 31 2018

Fig 5: Expression of CD5 on thymocytes, ELISA for Shh in E17.5 thymus, and detection of Hh pathway activation in E17.5 WT and Gli3 mutant thymocytes. (A-C) Flow cytometry analysis of E17.5 FTOC +4 days, with scatter plots showing relative mean±s.e.m., where each point represents thymus from an individual embryo from Gli3+/+ (WT; n=14, circles), Gli3+/- (n=14, squares), Gli3-/- (n=11, triangles). (A) CD5 staining on DP, SP4 and SP8 populations, showing MFIs from a representative experiment. (B) MFI of CD5 on SP4, DP and SP8 populations, giving significance by Student's t-test compared with WT for SP4 (Gli3+/-, P<0.007; Gli3-/-, P<0.004), DP (Gli3-/-, P<0.001) and SP8 (Gli3-/-, P<0.004). (C) MFI of CD5, gated on CD3hi on SP4, DP and SP8, giving significance by Student's t-test for CD3hi SP4 (Gli3-/-, P<0.03), CD3hi DP (Gli3-/-, P<0.04) and CD3hi SP8 (Gli3-/-, P<0.04). (D) Shh protein measured by ELISA in Gli3+/+ (n=3) and Gli3-/- (n=3) E17.5 thymus (P<0.003). (E-I) Flow cytometry analysis of GFP expression in SP4, DP and SP8 populations from GBS-GFP-tg E17.5 FTOC +4 days from Gli3+/+ (n=3), Gli3+/- (n=4) and Gli3-/- (n=3) littermates. Scatter plots show relative mean± s.e.m., where each point represents thymus from a different embryo. (E) SSC versus GFP, gated on SP4 (top row), DP (middle row) and SP8 (bottom row) cells from Gli3+/+ (left), Gli3+/- (middle) and Gli3-/- (right) littermates, with percentage in region shown. (F) Percentage GFP+, giving significance by Student's t-test compared with WT for SP4 (Gli3+/-, P<0.004), DP (Gli3-/-, P<0.003) and SP8 (Gli3+/-, P<0.02; Gli3-/-, P<0.02). (G) MFI of GFP, giving significance by Student's t-test compared with WT for SP4 (Gli3+/-, P<0.003; Gli3-/-, P<0.02), DP (Gli3-/-, P<0.04) and SP8 (Gli3-/-, P<0.004). (H) Percentage of GFP+ cells, gated on CD3hi, giving significance by Student's t-test compared with WT for CD3hi SP4 (Gli3+/-, P<0.006; Gli3-/-, P<0.01) and CD3hi SP8 (Gli3+/-, P<0.05). (I) MFI of GFP, gated on CD3hi, giving significance by Student's t-test compared with WT for SP4 (Gli3-/-, P<0.01) and SP8 (Gli3+/-, P<0.01; Gli3-/-, P<0.01).

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FITC anti-mouse CD5

BioLegend's FITC anti-mouse CD5 is a Rat monoclonal antibody. The FITC anti-mouse CD5 Antibody was generated using CD5, and CD5 Antigen as the antigen. It reacts with Mouse. This antibody has been shown to work in applications such as: Flow Cytometry.

Description

FITC anti-mouse CD5 [53-7.3]

Product Specs
ItemFITC anti-mouse CD5
CompanyBioLegend
Price Pricing Info Supplier Page View Company Product Page
Catalog Number100605
NCBI Gene AliasesLy-1, Ly-12, Ly-A, Lyt-1
Clone53-7.3
Quantity50 µg
ApplicationsFlow Cytometry (FC)
IsotypeIgG2a, Kappa
ReactivityMouse
Conjugate/TagFITC
HostRat
Antibody TypePrimary
NCBI Full Gene NameCD5 antigen
Concentration0.50 mg/mL
FormatPhosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
ImmunogenMouse thymus or spleen
ClonalityMonoclonal
TargetCD5

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4/16/2021CD5 Antibody Gives Great Results Preeyam PatelWe are interested in detection of B1a and B1b cells in mouse spleen.

Development (Cambridge, England)

Development (Cambridge, England)

Development (Cambridge, England)

Development (Cambridge, England)

Development (Cambridge, England)

FITC anti-mouse CD5 from BioLegend

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FITC anti-mouse CD5 from BioLegend

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