Journal: Cell Reports

Article Title: Psoriatic and rheumatoid arthritis joints differ in the composition of CD8+ tissue-resident memory T cell subsets

doi: 10.1016/j.celrep.2023.112514

Figure Lengend Snippet:

Article Snippet: Purified anti-Human IL-17F (LN2-9C4) Antibody (panel II; −173Yb) , Miltenyi Biotech , Custom.

Techniques: Purification, Recombinant, Staining, Blocking Assay, Amplification, Software

Journal: Clinical and Translational Medicine

Article Title: XBP1‐elicited environment by chemotherapy potentiates repopulation of tongue cancer cells by enhancing miR‐22/lncRNA/KAT6B‐dependent NF‐κB signalling

doi: 10.1002/ctm2.1166

Figure Lengend Snippet: Cytotoxic treatment‐induced dying cells promoted tumour cell repopulation via secretion of amphiregulin (AREG) and basic fibroblast growth factor (bFGF). (A) SCC‐25 cells were treated with or without cisplatin (cDDP) (8 µg/ml) for 24 h, and the conditioned medium (CM) was collected. The differentially expressed cytokines in the CM were identified using a cytokine array. (B) SCC‐25, SCC‐15 and SCC‐9 cells were treated with cDDP in combination with vitamin E (VitE), hydralazine (Hlz) or 4µ8cα for 24 h. The CM from further incubation for 24 h was collected. The protein levels of AREG and bFGF in CM were determined by enzyme‐linked immunosorbent assay (ELISA) ( n = 3), ** p < .01, *** p < .001. (C and D) SCC‐25, SCC‐15 and SCC‐9 cells were treated with cDDP for 24 h. Another new SCC‐25, SCC‐15 and SCC‐9 cells were cocultured with feeder cells combination with AREG‐neutralising antibody (1 µg/ml), bFGF‐neutralising antibody (1 µg/ml) and Akt inhibitor MK2206 (800 µM). Transactivation of NF‐κB signalling in mentioned cells was measured by Cignal Reporter Assays ( n = 3), ** p < .01, *** p < .001, **** p < .0001 (C). p‐Akt, Akt, p‐IκBα, IκBα and β‐Actin were detected by western blots (D). (E) SCC‐25, SCC‐15 and SCC‐9 feeder cells were treated with cDDP for 24 h. SCC‐25/Luc, SCC‐15/Luc and SCC‐9/Luc reporter cells were cocultured with feeder cells in combination with neutralising antibody against AREG or bFGF or Akt inhibitor MK2206 in 24‐well plates. Cancer cell repopulation in vitro was observed by luciferase activities, ** p < .01. (F) SCC‐25 cells were treated with cDDP for 24 h. SCC‐25/Luc cells were injected subcutaneously together with cDDP‐treated SCC‐25 cells in nude mice to generate xenograft tumours. The mice were randomly grouped and received i.p. injection of MK2206 (300 mg/kg) or phosphate‐buffered saline (PBS) once every 3 days, and tumour growth was represented by luciferase levels, *** p < .001. Data in (B), (C), (E) and (F) are represented as mean ± SEM. Statistical significance was determined by a two‐tailed Student's t ‐test

Article Snippet: Primary antibodies (Abs) against amphiregulin (AREG) (AB‐262‐NA) and basic fibroblast growth factor (bFGF) (AF‐233‐NA) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Western Blot, In Vitro, Luciferase, Injection, Saline, Two Tailed Test

Journal: Clinical and Translational Medicine

Article Title: XBP1‐elicited environment by chemotherapy potentiates repopulation of tongue cancer cells by enhancing miR‐22/lncRNA/KAT6B‐dependent NF‐κB signalling

doi: 10.1002/ctm2.1166

Figure Lengend Snippet: Molecular events for tumour cell repopulation correlate with poor clinical outcome. (A) The levels of X‐box binding protein‐1 (XBP1), amphiregulin (AREG) and basic fibroblast growth factor (bFGF) in representative cancer specimens were examined by immunohistochemistry (IHC) assays. Scale bar = 50 µm. (B) The correlation between the expression levels of XBP1s and AREG or bFGF was analysed. (C) The average IHC values of each protein (XBP1s, AREG, bFGF) in 88 tongue cancer tissues were obtained, and IHC scores above the average value indicated high expression, otherwise indicated low expression. Kaplan–Meier analysis indicated a correlation between XBP1s/AREG/bFGF levels and overall survival in patients with tongue cancer. Kaplan–Meier survival analysis is shown here with a two‐sided log‐rank p ‐value. (D) The protein levels of AREG and bFGF in serum were examined by enzyme‐linked immunosorbent assay (ELISA) ( n = 3), ** p < .01, *** p < .001. Data are presented as the mean value ± SEM. Statistical significance was determined by a two‐tailed Student's t ‐test. (E) The correlation between the XBP1s/AREG/bFGF levels and tongue cancer recurrence was analysed. (F) Fifteen paired tissues of primary tongue cancer and recurrent tongue cancer were collected. The protein levels of p65, IκBα and lysine acetyltransferase 6B (KAT6B) were examined by IHC assays. The transcript levels of miR‐22‐3p, XLOC_003973 and XLOC_010383 were examined by in situ hybridisation (ISH) assays. Scale bars = 50 µm

Article Snippet: Primary antibodies (Abs) against amphiregulin (AREG) (AB‐262‐NA) and basic fibroblast growth factor (bFGF) (AF‐233‐NA) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay, Immunohistochemistry, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test, In Situ, Hybridization

Journal: Neuron

Article Title: IGF1-dependent synaptic plasticity of mitral cells encodes olfactory memory during social learning

doi: 10.1016/j.neuron.2017.06.015

Figure Lengend Snippet: (A) Coronal section of the MOB (left) and cropped region (right) showing layer-specific expression of Syt1 (red) in WT mice. (B) Example micrographs from the EPL showing localizations of Syt1 puncta (red) relative to vGlut1+ (green) and vGAT+ (blue) synapses. (C, D) Coronal sections of the MOB (left) and cropped region (right) showing layer-specific expression of Syt10 (green) in WT mice (C) and in Syt10 KO mice (D). (E, F) Example micrographs from the EPL showing localizations of Syt10 puncta (green) relative to Syt1+ (red) and vGlut1+ (blue) synapses (E), and Syt10 puncta (green) relative to vGlut1+ (red) and vGAT+ (blue) synapses (F). For more data of synaptic localization of Syt1 and Syt10, see Figure S4 and S5. (G) Example G-M IPSC traces (left), summarized input-output curves (middle) and curve slopes (right) from M72-associated mitral cells in control (Ctrl) and Syt1 KD MOB slices of naïve M72-GFP mice. For Syt1 KD efficiency and OS-M EPSC data, see Figure S6 (A–E). (H) Example G-M IPSC traces (left), summarized input-output curves (middle) and curve slopes (right) from M72-associated mitral cells in control (Ctrl) and Syt10 KO (Cre) MOB slices of naïve M72-GFP + Syt10f/f mice. For Syt10 KO efficiency, see Figure S6 (F–H). (I) Schematic diagram showing AAV injection, STFP training and subsequent slice recordings of G-M IPSCs from M72-associated mitral cells in M72-GFP + Syt10 f/f mice. (J–L) Example G-M IPSC traces (left), summarized input-output curves (middle) and curve slopes (right) from M72-associated mitral cells in control (Ctrl) and Syt10 KO (Cre) MOB of M72-GFP + Syt10 f/f observers of plain food (J), Oct-food (K) and Acp-food (L). Data in (G, H, J, K, L) are means ± SEM (error bars), with mouse number as sample size n. Numbers of neurons/mice are indicated in bars. Statistical analysis is by Student’s t test (***P < 0.001; *P < 0.05; n.s. P > 0.1).

Article Snippet: Syt10 , NeuroMab , Cat. No. 75-262; RRID: AB_10671950.

Techniques: Expressing, Control, Injection

Journal: Neuron

Article Title: IGF1-dependent synaptic plasticity of mitral cells encodes olfactory memory during social learning

doi: 10.1016/j.neuron.2017.06.015

Figure Lengend Snippet: (A) Schematic diagram showing AAV injection, food deprivation and subsequent food-finding test. (B–D) Analyses of time spent in finding buried food by the adult mice with Syt1 KD (B), Syt10 KO (C) and IGF1-R KO (D) in the MOB. (E) Schematic diagram showing AAV injection, habituation, STFP training and memory tests after STFP. (F–H) Analyses of memory retrieval for Acp-flavored food at different time points (immediately, 1 day and 14 days) after STFP by adult mice with intrabulbar Syt1 KD (F), Syt10 KO (G) and IGF1-R KO (H). For other behavioral tests, see Figure S7. Data (B–D, F–H) are means ± SEM (error bars), with mouse number as sample size n. Numbers of mice are indicated in bars. Statistical analysis is by Student’s t test (***P < 0.001; **P < 0.01; *P < 0.05; n.s. P > 0.1).

Article Snippet: Syt10 , NeuroMab , Cat. No. 75-262; RRID: AB_10671950.

Techniques: Injection

Journal: Neuron

Article Title: IGF1-dependent synaptic plasticity of mitral cells encodes olfactory memory during social learning

doi: 10.1016/j.neuron.2017.06.015

Figure Lengend Snippet: (A) Odor exposure without a social context induces transient Ca2+ entry that triggers Syt1-dependent glutamate release but not Syt10-dependent IGF1 secretion from mitral cell dendrite. Such process then evokes GABA release from granule cell and feedback inhibition on the mitral cell without G-M synaptic potentiation. (B) Odor exposure in a social context during STFP induces sustained Ca2+ entry in mitral cell dendrites and triggers both glutamate release and IGF1 exocytosis. IGF1 acts on IGF1-R and potentiates G-M synaptic strength by recruiting more GABA-R, which may be a synaptic substrate for the association of odor signals and social context.

Article Snippet: Syt10 , NeuroMab , Cat. No. 75-262; RRID: AB_10671950.

Techniques: Inhibition

Journal: Neuron

Article Title: IGF1-dependent synaptic plasticity of mitral cells encodes olfactory memory during social learning

doi: 10.1016/j.neuron.2017.06.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Syt10 , NeuroMab , Cat. No. 75-262; RRID: AB_10671950.

Techniques: Plasmid Preparation, Patch Clamp, Imaging, Software

Journal: PeerJ

Article Title: Four tyrosine residues of the rice immune receptor XA21 are not required for interaction with the co-receptor OsSERK2 or resistance to Xanthomonas oryzae pv. oryzae

doi: 10.7717/peerj.6074

Figure Lengend Snippet: (A) Representation of the XA21 juxtamembrane (JM), kinase subdomains (I–XI), and C-terminal region (CT) (XA21JK). Full-length XA21 has N-terminal leucine-rich repeats and a transmembrane domain that are not depicted. The positions of the nine XA21JK tyrosine residues (Y) are indicated in black. The conserved lysine (K) required for kinase activity and aspartate (D) required for phospho-transfer are labeled in orange. Previously studied XA21 JM Threonine (T) and Serine (S) residues are depicted in blue. (B) Immunoblots of wildtype (WT) GST-tagged XA21 (GST-XA21JK) and GST-XA21JK with lysine modified to glutamine (GST-XA21JK K736E ) and WT GST-XA21JK and variants with individual tyrosine residues modified to phenylalanine (Y698F, Y723F, Y786F, Y829F, Y889F, Y894F, Y907F, Y909F, Y936F). Tyrosine-specific phosphorylation was detected by immunoblotting with anti-phosphotyrosine antibodies (anti-pY). Equal loading of proteins was confirmed by immunoblotting with an anti-GST antibody and/or Coomassie Brilliant Blue (CBB) staining of the membrane. Densitometry measurements shown below the panel represent the ratio between phosphotyrosine and GST signals normalized to WT GST-XA21JK. The above experiments were performed twice with similar results.

Article Snippet: Detection of anti-GST and anti-phosphotyrosine primary antibodies was performed using an Alexa Fluor 680-conjugated secondary antibody (Rockland Immunochemicals, Gilbertsville, PA, USA), and imaged using an Odyssey® Infrared (LI-COR Biosciences, Lincoln, NE, USA) imaging system.

Techniques: Activity Assay, Labeling, Western Blot, Modification, Phospho-proteomics, Staining, Membrane

Journal: PLoS Pathogens

Article Title: Ebola virus-mediated T-lymphocyte depletion is the result of an abortive infection

doi: 10.1371/journal.ppat.1008068

Figure Lengend Snippet: ( A-E ) The number of viral genomic RNA (A) and specific viral mRNAs (B-E) copies/ng in CD4 + T-cells exposed to EBOV for 1, 2 and 5 days, determined by DDRT-PCR with background signals in mock-infected cells subtracted. ( F ) Histograms displaying the relative number of sequence reads for each viral mRNA at day 1 and 4 as determined by deep sequencing. Bottom right: the zoomed in fragment of the reads corresponding to the 5’-terminal part of the genome, which includes the L gene. ( G ) Numbers of viral genomic RNA (gRNA, left) and mRNA (right) copies normalized to GAPDH in mock-infected, EBOV-infected and 1E7-03 treated EBOV-infected cells. ( H ) Ratios of viral gRNA to mRNA in non-treated and 1E7-03 treated cells. ( I ). Analysis of the total and phosphorylated VP30 in Jurkat and SUP-T1 T cells and Huh7 cells incubated with EBOV for 2 or 5 days at MOI 3 PFU/cell by Western blot. Panel A: 1 of 3 independent experiments. Panels B-E: Average of duplicate reads from EBOV-exposed CD4 + T-cells isolated from one of 3 donors. Panel F: Representative number of reads from deep sequencing results depicted as histogram shown from 1 of 4 independent donors. Panel G: data from triplicate samples from one representative donor; two donors were analyzed resulting in similar data. *** P<0.001, **** P<0.0001.

Article Snippet: Antibodies raised against phosphorylated EBOV VP30 S29-31 (phS29-31) were custom ordered from ProSci.

Techniques: Infection, Sequencing, Incubation, Western Blot, Isolation