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: PLoS ONE

Article Title: Acetylcholine Acts on Androgen Receptor to Promote the Migration and Invasion but Inhibit the Apoptosis of Human Hepatocarcinoma

doi: 10.1371/journal.pone.0061678

Figure Lengend Snippet: (A–D) Western blots showed the effects of Ach and MEC on STAT3 phosphorylation (A, B) and Akt phosphorylation (C, D) and in SNU-449 cells. Phosphorylation of STAT3 (A) and Akt (C) was enhanced by Ach in SNU-449 cells, whereas acetylcholine receptor antagonists MEC blocked the effect of Ach on STAT3 (B) and AKT (D) phosphorylation.

Article Snippet: Primary antibodies used here included rabbit monoclonal anti-AR (D6F11) (3H8; Cell Signaling Technology, Danvers, MA), rabbit monoclonal anti-nicotinic acetylcholine receptor alpha 7 (ab10096, abcam, USA), rabbit monoclonal anti-pSTAT3, anti-pAKT (195-14; BioCheck, Foster City, CA), and monoclonal anti-β-actin (Sigma).

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Stimulus-Specific Expression, Selective Generation and Novel Function of Grass Carp ( Ctenopharyngodon idella ) IL-12 Isoforms: New Insights Into the Heterodimeric Cytokines in Teleosts

doi: 10.3389/fimmu.2021.734535

Figure Lengend Snippet: Signaling pathways of rgcIl-12 isoforms in regulating il-17a/f1 in lymphocytes. (A) Lymphocytes were incubated with rgcIl-12AB (1000 ng/mL) in the presence or absence of STAT3 VI (STAT3 inhibitor, 30 µM). (B) Lymphocytes were incubated with RgcIl-12AB (1000 ng/mL) in the presence or absence of SR1001 (RORγt inhibitor, 15 µM). The il-17a/f1 mRNA was normalized by β-actin and expressed as fold changes compared with the control group. (C) Phosphorylation of Stat3 was detected in the lymphocytes treated with rgcIl-12AB (1000 ng/mL) for 0-60 min. (D) Lymphocytes were incubated with rgcIl-12BB (1000 ng/mL) in the presence or absence of STAT3 VI (STAT3 inhibitor, 30 µM). (E) Lymphocytes were incubated with rgcIl-12BB (1000 ng/mL) in the presence or absence of SR1001 (RORγt inhibitor, 15 µM). The il-17a/f1 mRNA was normalized by β-actin and expressed as fold changes compared with the control group. Data are shown as mean ± SEM (N = 4). Different letters indicate significant differences at p < 0.05. (F) Phosphorylation of Stat3 was detected in lymphocytes treated with rgcIl-12BB (1000 ng/mL) for 0-60 min.

Article Snippet: To detect the activation of signaling pathways, the cell lysates were separated on 12% SDS-PAGE and the signaling molecules were detected by WB assay by using specific primary antibodies for phosphorylated ERK (anti-pERK1/2, 1:1000, Cell Signaling Technology), JNK (anti-pJNK, 1:1000, Cell Signaling Technology), p38 (anti-p-p38, 1:1000, Cell Signaling Technology), IκBa (anti-p-IκBa, 1:1000, Cell Signaling Technology), STAT3 (anti-pSTAT3, 1:1000, Anaspec, Fremont, CA, USA), and the β-actin (anti-β-actin 1:5000, Boster, Wuhan, China) as the loading control.

Techniques: Incubation

Journal: Frontiers in Immunology

Article Title: Stimulus-Specific Expression, Selective Generation and Novel Function of Grass Carp ( Ctenopharyngodon idella ) IL-12 Isoforms: New Insights Into the Heterodimeric Cytokines in Teleosts

doi: 10.3389/fimmu.2021.734535

Figure Lengend Snippet: The effects of gcTgf-β1 on rgcIl-12AB- and rgcIl-12BB-induced il-17a/f1 and ifn-γ mRNA expression. Lymphocytes were treated with 1000 ng/mL of rgcIl-12AB (A, B) and rgcIl-12BB (C, D) in the presence or absence of rgcTgf-β1 (100 ng/mL) for 12 h. (E, F) Lymphocytes were exposed to rgcIl-12BB (1000 ng/mL) in the presence or absence of gcTgf-β1 mAb (1:1000 diluted) for 12 h. (G, H) Lymphocytes were incubated with rgcIl-12BB (1000 ng/mL) in the presence or absence of ALK5 inhibitor (TGF-β1 RI Kinase inhibitor VIII, 2 µM) for 12 h. Then the levels of il-17a/f1 and ifn-γ mRNA were detected by qPCR. Relative mRNA levels were analyzed using β-actin as an internal reference and expressed as the fold changes of the control group (blank column). Data are shown as mean ± SEM (N = 4). Different letters indicate significant differences at p < 0.05. (I) Phosphorylation of Stat3 was detected in lymphocytes incubated with rgcIl-12BB (1000 ng/mL) in the presence or absence of rgcTgf-β1 (100 ng/mL) for 20 min.

Article Snippet: To detect the activation of signaling pathways, the cell lysates were separated on 12% SDS-PAGE and the signaling molecules were detected by WB assay by using specific primary antibodies for phosphorylated ERK (anti-pERK1/2, 1:1000, Cell Signaling Technology), JNK (anti-pJNK, 1:1000, Cell Signaling Technology), p38 (anti-p-p38, 1:1000, Cell Signaling Technology), IκBa (anti-p-IκBa, 1:1000, Cell Signaling Technology), STAT3 (anti-pSTAT3, 1:1000, Anaspec, Fremont, CA, USA), and the β-actin (anti-β-actin 1:5000, Boster, Wuhan, China) as the loading control.

Techniques: Expressing, Incubation