Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Journal of Cellular Signaling

Article Title: Phorbol-12-Myristate-13-Acetate (PMA) Reactivates Replication from HIV-1 Latency and Induces Jurkat Cell Death

doi: 10.33696/signaling.5.110

Figure Lengend Snippet: Figure 3. The effects of AZT interruption on P-TEFb pathways and Jak/Stat pathways in HIV-1 infected Jurkat cells. As illustrated in Study Design I, shown in Supplementary Figure 2A, Jurkat cells were exposed to HIV-1 (MN stained) for 2 hours. Subsequently, the cells were washed twice with PBS and cultured at 37°C for 7 days. On day 7, some HIV-1-infected cells were transferred to fresh medium and cultured for an additional 7 days (Treatment A), while others were treated with AZT (2 µm) for 7 days (Treatment B & C). On day 14, equal amounts of live cells were transferred to fresh medium and cultured for another 7 days: Medium only (Treatment A); AZT (2 µm) continuously (Treatment B); AZT interruption (Treatment C). To examine if PMA can activate p-TEFb pathways and Jak/Stat pathways, on day 21, total cell lysates were subjected to Western blot analysis to detect the expression of proteins that crucial components of the signaling pathways associated with p-TEFb activation, (A) Brd4, CDK9 and Cyclin T1; (B) P-RPb-CTD; and (C) Jak2 and Atat3.

Article Snippet: The membranes were then incubated with primary antibodies, specifically rabbit polyclonal/mouse monoclonal antibodies targeting AKT, ATG5, Beclin1, Brd4, CD3ζ, CD4, CD28, CDK9, FADD, Fas, FLIP, Jak2, p53, PI3K, PLCγ1, SP1, ULK1, and GAPDH, which were sourced from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Infection, Staining, Cell Culture, Western Blot, Expressing, Protein-Protein interactions, Activation Assay

Journal: iScience

Article Title: The mutation of NSUN5 R295C promotes preeclampsia by impairing decidualization through downregulating IL-11Rα

doi: 10.1016/j.isci.2024.108899

Figure Lengend Snippet: The mutation of Nsun5 suppresses JAK/STAT signal pathway and downregulates Cyclin D3 (A) QPCR results showing the expressions of Jak2 , Stat3 , Cyclin D3 , and P21 in the decidua of NSUN5 R295C and WT mice at E7.5. (B) The expressions of JAK2, P-JAK2, STAT3, P-STAT3, and Cyclin D3 analyzed by western blot (n = 6 per genotype). The protein levels of P-JAK2, P-STAT3, and Cyclin D3 were decreased in the decidua of NSUN5 R295C mice when compared to those in WT mice. (C–E) The results of western blotting were quantified and shown in bar graphs. (C) The bar graphs show the relative density of P-JAK2 and JAK2 according to the statistical analysis of the results of three independently repeated experiments. (D) The bar graphs show the relative density of P-STAT3 and STAT3 according to the statistical analysis of the results of three independently repeated experiments. (E) The bar graph shows the relative density of Cyclin D3 according to statistical analysis of the results of three independently repeated experiments. (F and G) NSUN5 binds to Il-11rα mRNA in decidua as revealed by RNA immunoprecipitation (RIP). (H) Schematic diagram showing the mechanisms of NSUN5-IL-11Rα interaction. NSUN5 mutation leads to IL-11Rα down expression. Insufficient phosphorylation of JAK2 and STAT3 downregulates Cyclin D3, resulting in decreased polyploidization, and eventually leading to PE development. Data are represented as mean ± SEM analyzed using Student’s t test. ∗p < 0.05; ∗∗p < 0.01.

Article Snippet: Phospho-JAK2-Y1007 Rabbit pAb , ABclonal , Cat. NO: Ap0917; RRID: AB_2863830.

Techniques: Mutagenesis, Western Blot, Immunoprecipitation, Expressing

Journal: iScience

Article Title: The mutation of NSUN5 R295C promotes preeclampsia by impairing decidualization through downregulating IL-11Rα

doi: 10.1016/j.isci.2024.108899

Figure Lengend Snippet:

Article Snippet: Phospho-JAK2-Y1007 Rabbit pAb , ABclonal , Cat. NO: Ap0917; RRID: AB_2863830.

Techniques: Enzyme-linked Immunosorbent Assay, Immunoprecipitation, Software