3168002b  (fluidigm)

Journal: Frontiers in Pharmacology

Article Title: Compensatory attenuation of cortical apoptosis by SK2 downregulation following ketamine anesthesia

doi: 10.3389/fphar.2026.1761187

Figure Lengend Snippet: Apamin-sensitive SK2 channel-mediated mAHP currents may be linked to changes in neuronal spike frequency and adaptation at 24 h post-ketamine anesthesia. (A,B) Representative traces (A) and amplitudes (B) of the mAHP currents, treatment conditions as indicated. S1 slices from control (Ctrl) and ketamine-treated (Ket) rats were separately incubated and perfused with apamin (100 nM) or its vehicle. 22–25 neurons from 7–10 rats were used per condition. (C) Plots of spike frequency vs. current injected for layer II/III pyramidal neurons of S1. The significant differences in the spike frequency between the Ctrl and Ket groups (Ctrl: Vehicle vs. Ket: Vehicle, 80 pA, 100 pA and 110 pA, P < 0.01; 90 pA, P < 0.001) were eliminated after apamin treatment (Ctrl: Apamin vs. Ket: Apamin, P > 0.05). 20–22 neurons from 7–9 rats were recorded per condition. (D,E) Spikes in S1 layer II/III pyramidal neurons evoked for 3 s, 80 pA current injection (D) , and the adaptation index (E) was obtained by the algorithm mentioned above. 18–20 neurons from 7–9 rats were recorded per condition. (F) Quantitative analysis of SK1-3 mRNA in S1 of P8 rats. 5 rats were used per condition. P > 0.05. (G,H) Immunoblots and quantitative analysis of total (G) and membrane-bound (H) SK1-3 levels in S1 of ketamine-treated rats, normalized to corresponding levels in control rats. 8–12 rats were used per condition. * P < 0.05, ** P < 0.01, *** P < 0.001; n.s ., not significant. Data were analyzed using the Mann-Whitney U test for (F,H) and unpaired two-tailed Student’s t-tests for the other panels. Data are shown as the mean ± SEM.

Article Snippet: The following primary antibodies were used: SK1 (1:500, Alomone Labs, Cat# APC-039), SK2 (1:500, Alomone Labs, Cat# APC-028), SK3 (1:500, Alomone Labs, Cat# APC-025), β-actin (1:1,000, Millipore, Cat# A1978 ), and pan-cadherin (1:1,000, Sigma-Aldrich, Cat# SAB4500001 ).

Techniques: Control, Incubation, Injection, Western Blot, Membrane, MANN-WHITNEY, Two Tailed Test

Journal: Nature Immunology

Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

doi: 10.1038/s41590-025-02381-7

Figure Lengend Snippet: a , sjTREC quantification in HDs (black dots), heterozygous donors (gray dots) and individuals with CBL -LOH (colored dots), as determined by qPCR of whole-blood DNA; WBCs, white blood cells. b , Recent thymic emigrant CD4 + and CD8 + T cells quantified in peripheral fresh blood by mass cytometry and gating of CD31 + cells among naive T cells; data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. c , Quantification of the indicated T cell subsets in the peripheral blood of HDs, heterozygous HDs and individuals with CBL -LOH of the indicated ages as determined by mass cytometry; data are shown as mean ± s.d. The statistical significance of differences was assessed by multiple two-sided Mann–Whitney tests, with correction for multiple testing; * P < 0.05 and ** P < 0.005. In b and c , controls 0–3 years old ( n = 2), controls 4–15 years old ( n = 9), controls 16–100 years old ( n = 28), pediatric participants (LOH) ( n = 5), adult participants (LOH) ( n = 2) and heterozygous individuals ( n = 3). d , Percentage of dead cells in cultures of activated fresh PBMCs from HDs, the heterozygous father and participants (P1–P3) after 5 days of TCR stimulation, as determined by dead cell marker staining and flow cytometry; n = 3 HDs and n = 3 patient; data are shown as mean ± s.d. e , f , Cell division index of CD4 + ( e ) and CD8 + ( f ) T cells of HDs, the heterozygous father and participants (P1–P3) after 5 days of the indicated TCR stimulation, as determined by dilution of CFSE; n = 3 HDs and n = 3 patients; data are shown as mean ± s.d. g , Cytokine response by STAT5 phosphorylation (left) and cytokine production (right) by T cell blasts that are homozygous (red), heterozygous (gray) and homozygous WT for CBL Ub LOF variants following the indicated stimuli. The bars show the mean of the displayed data points (one for each T blast line); NS, not significant; TEMRA, terminally differentiated effector memory T cells; MAIT, mucosal-associated invariant T cells; MFI, median fluorescence intensity; PHA, phytohemagglutinin; PMA, phorbol 12-myristate 13-acetate.

Article Snippet: 168Er , CD8 , SK1 , 3168002B , Fluidigm.

Techniques: Mass Cytometry, MANN-WHITNEY, Marker, Staining, Flow Cytometry, Phospho-proteomics, Fluorescence

Journal: Nature Immunology

Article Title: Somatic deficiency of the human E3 ubiquitin ligase CBL in leukocytes impairs B cell but not T cell development and function

doi: 10.1038/s41590-025-02381-7

Figure Lengend Snippet: ( a ) CytoF immunophenotyping of patients total CD3 + T cells, NK cells and indicated NK cell subsets. Controls 0–3 y.o. (n = 2), Controls 4–15 y.o. (n = 9), Controls 16–100 y.o. (n = 28), pediatric patients (LOH) (n = 5), adult patients (LOH) (n = 2), heterozygous individuals (n = 3), mean ± s.d. Statistical significance was assessed with multiple Mann Whitney tests adjusted for multiple testing. *p<0.05. ( b ) Assessment of T cell proliferation of patients P1, P2 and P3. (top) gating strategy, (bottom) CFSE dilution plots. ( c ) Intracellular TNF production of CD4 + and CD8 + T cell blasts in homozygous (n = 4), heterozygous (n = 2) and wildtype (n = 5) state of CBL Ub LOF variants. Line indicates the mean of the displayed datapoints. ( d,e ) Intracellular and extracellular cytokine production by patients T fh cells stimulated with CD2/CD3/CD28 under T h 0 conditions for 5 days. ( d ) Intracellular staining of the indicated cytokines. Healthy donors (n = 17), CBL-LOH patients (n = 4). Mann-Whitney testing with correction for multiple testing did not reveal significant (p< 0.05) differences between healthy donors and CBL-LOH patients. Line indicates the mean of the displayed datapoints. ( e ) Extracellular detection of cytokines by ELISA. Healthy donors (n = 6), CBL-LOH patients (n = 3). Mann-Whitney testing with correction for multiple testing did not reveal significant (p< 0.05) differences between healthy donors and CBL-LOH patients. Line indicates the mean of the displayed datapoints.

Article Snippet: 168Er , CD8 , SK1 , 3168002B , Fluidigm.

Techniques: MANN-WHITNEY, Staining, Enzyme-linked Immunosorbent Assay