pcreb 1 s133  (Santa Cruz Biotechnology)

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Mass Cytometry Assessment of Cell Phenotypes and Signaling States in Human Whole Blood

doi: 10.1007/978-1-0716-2553-8_10

Figure Lengend Snippet: Whole blood phosphoflow panel 1

Article Snippet: 142 Nd cCasp3 D3E9 Fluidigm 3142004A 143 Nd CD19 HIB19 Biolegend 302202 144 Nd pPLCg2 [Y759] K86-689.37 Fluidigm 3144015A 145 Nd CD4 RPA-T4 Fluidigm 3145001B 146 Nd IgD IA6-2 Fluidigm 3146005B 147 Sm CD20 2H7 Fluidigm 3147001B 148 Nd IgA Polyclonal Fluidigm 3148007B 149 Sm CD25 2A3 Fluidigm 3149010B 150 Nd pStat5 [Y694] 47 Fluidigm 3150005A 151 Eu CD123 6H6 Fluidigm 3151001B 153 Eu pStat1 [Y701] 4a Fluidigm 3153005A 154 Sm CD45 HI30 Fluidigm 3154001B 155Gd CD27 L128 Fluidigm 3155001B 156 Gd p38 [T180/Y182] D3F9 Fluidigm 3156002A 157 Gd CD24 ML-5 Biolegend 311102 158 Gd pStat3 [Y705] 4/P-Stat3 Fluidigm 3158005A 159Tb CD11c Bu15 Fluidigm 3159001B 160Gd CD14 M5E2 Fluidigm 3160001B 161 Dy CD141(BDCA-3) AD5-14H12 Miltenyi 130-090-694 162 Dy CD66b 80H3 Fluidigm 3162023B 163 Dy CD56 NCAM16.2 Fluidigm 3163007B 164 Dy IkBa L35A5 Fluidigm 3164004A 165 Ho pCREB [S133] 87G3 Fluidigm 3165009A 166 Er CD16 B73.1 Biolegend 360702 167 Er CD38 HIT2 Fluidigm 3167001B 168 Er CD8 SK1 Fluidigm 3168002B 169 Tm CD45RA HI100 Fluidigm 3169008B 170 Er CD3 UCHT1 Fluidigm 3170001B 171 Yb pERK1/2 [T202/Y204] D13.14.4E Fluidigm 3171010A 172 Yb Anti-Ki-67 B56 Fluidigm 3172024B 174 Yb HLA-DR L243 Fluidigm 3174001B 175Lu CD7 CD7-6B7 Biolegend 343102 176 Yb CD127/IL-7Ra P48-48 Novus Bio MAB306-100 209Bi CD11b/Mac-1 ICRF44 Fluidigm 3209003B Open in a separate window 1 Open channels are not shown but include Pd channels, Cd channel, Pt channels, 89Y,152Sm and 173Yb Whole blood phosphoflow panel1.

Techniques:

Journal: bioRxiv

Article Title: SCA-1 micro-heterogeneity in the fate decision of dystrophic fibro/adipogenic progenitors

doi: 10.1101/2020.05.14.096438

Figure Lengend Snippet: (A) Experimental design to induce adipogenic differentiation of mdx FAP cell states. GM = growth medium; AIM = adipogenic induction medium; MM = maintenance medium. (B) and (C) Bar plots showing the percentage of ORO positive cells and PPAR-gamma positive cells per field. AM = adipogenic medium (AM = AIM + MM). n = 4. Statistical analysis was performed by a Two-way ANOVA. (D) Representative micrographs of (B) and (C). Cells were immunolabelled for PPAR-gamma (yellow) and nuclei were counterstained with Hoechst 33342 (blue). Lipid droplets were stained with ORO (red). (E) Experimental design applied to obtain fully differentiated adipocytes. (F) Bar plot indicating the percentage of ORO positive cells. n = 3. Statistical significance was calculated through a Student t-test. (G) Representative micrographs of (F). Lipid droplets in red (ORO staining) and nuclei in blue (Hoechst 33342). (H) Representative viSNE maps showing the expression of SCA-1 and phospho-CREB-1 assessed by mass cytometry in FAPs isolated from mdx mouse by MACS and cultured 72 hours either in GM or AIM. (I) Cluster 1 (in blue) and cluster 2 (in orange) obtained applying the FlowSOM algorithm and then mapped onto viSNE maps. (J) Plot representing the expression in arbitrary units of SCA-1 and phospho-CREB-1 identified in (J). (H) Representative viSNE maps of FAPs isolated by MACS from mdx mice and then incubated either to GM or to AIM. The upper maps show expression level of SCA-1, while the lower maps show the level of phospho-CREB-1. n = 3. (I) Representative viSNE maps showing the two clusters produced by the clustering approach. (J) Dot plot showing the expression level of SCA-1 and the level of phospho-CREB-1 (both in arbitrary units) in the metaclusters from (I). Data are presented as mean ± SEM. ** p ≤ 0.01, *** p ≤ 0.001. Scale bars 100 μm.

Article Snippet: The full list of antibodies purchased from Fluidigm is: anti-CD45 89Y (3089005B); anti-CD146 141Pr (3155006B); anti-cleaved CASP-3 142Nd (3142004A); anti-CD34 144Nd (3143009B); anti-phospho-EGFR 146Nd (3146007A); anti-CD140a 148Nd (3148018B); anti-pRB 150Nd (3150013A); anti-CD140b 151Eu (3151017B); anti-Vimentin 154Sm (3154014A); anti-CD90.2 156Gd (3156006B); anti-phospho-STAT3 158Gd (3158005A); anti-CXCR-4 159Tb (3159030B); anti-Integrin alpha-7 161Dy (67-0010-05); anti-SCA-1 164Dy (3164005B); anti-phospho-CREB-1 176Yb (3176005A); DNA1 191Ir and DNA2 193Ir (201192A); Cisplatin 195Pt (201194); anti-CD31 165Ho (3165013B); anti-Actin 175Lu (3175026A); anti-c-kit 166Er (3166004B).

Techniques: Staining, Expressing, Mass Cytometry, Isolation, Cell Culture, Incubation, Produced