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: Archives of Medical Science : AMS

Article Title: Inhibitory effect of PDGF-BB and serum-stimulated responses in vascular smooth muscle cell proliferation by hinokitiol via up-regulation of p21 and p53

doi: 10.5114/aoms.2018.75085

Figure Lengend Snippet: Effect of hinokitiol on PDGF-BB signaling pathway in VSMCs. A – Confluent cells were pre-treated in the presence or absence of hinokitiol (1–100 μM) in a serum-free medium, and then stimulated with 10 ng/ml PDGF-BB for 10 min. The cells were lysed, and the levels of ( A ) p-ERK1/2, ( B ) p-Akt, ( C ) p-PI3K and ( D ) p-JAK2 were analyzed by SDS-PAGE and immunoblotting. The respective total proteins and a-tubulin were used for the normalization of phosphorylated ones. Data are expressed as mean ± SEM ( n = 3). * p < 0.05 compared to the PDGF-BB stimulated cells

Article Snippet: Anti-phospho-ERK1/2 (Thr 202 /Tyr 204 ), anti-phospho-AKT (Ser 473 ), antiphospho-PI3K, and antiphospho-JAK2 monoclonal antibodies (mAbs) were purchased from Cell Signaling (Beverly, MA, USA).

Techniques: SDS Page, Western Blot

Journal: Scientific Reports

Article Title: LncTUG1 promotes hepatocellular carcinoma immune evasion via upregulating PD-L1 expression

doi: 10.1038/s41598-023-42948-8

Figure Lengend Snippet: TUG1 affects the JAK2/ STAT3 pathway to regulate PD-L1. ( A ): The relative mRNA expression of JAK2 was analyzed in shTUG1 HCC cells. ( B ): The relative mRNA expression of STAT3 was analyzed in shTUG1 HCC cells. ( C ): The protein levels of pJAK2, JAK2, pSTAT3, STAT3, PD-L1 were analyzed in shTUG1 HCC cells by western blotting. ( D ) pJAK2, JAK2,and PDL1 protein levels are assessed by immunoblotting in HCC-LM3 cell was transfected with TUG1 silenced (shTUG1) and/or JAK2 overexpressed (JAK2-OE). ( E ) pSTAT3,STAT3,and PDL1 protein levels are assessed by immunoblotting in HCC-LM3 cell was transfected with TUG1 silenced (shTUG1) and/or STAT3 overexpressed (STAT3-OE).

Article Snippet: Protein detection was performed by using the following primary antibodies: anti-PD-L1 (ab205921, Abcam), anti-GAPDH (ab8245, Abcam), anti-JAK2 (#3230, Cell Signaling Technology), anti-pJAK2 Tyr1007/1008 (#3771,Cell Signaling Technology),anti-STAT3 (#9139, Cell Signaling Technology), anti-pSTAT3 Tyr705 (#9145, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Transfection

Journal: Discover oncology

Article Title: Hepatoma cell-derived exosomal SNORD52 mediates M2 macrophage polarization by activating the JAK2/STAT6 pathway.

doi: 10.1007/s12672-024-01700-y

Figure Lengend Snippet: Fig. 3 SNORD52 mediates M2 macrophage polariza- tion and activates JAK2/ STAT6 pathway. A Following transfection of OE-SNORD52/ OE-NC in THP-1 and U937 for 48 h, the mRNA expression of SNORD52 was detected by qRT-PCR. ****P < 0.0001 vs. OE-NC. B Following trans- fection of OE-SNORD52/ OE-NC in THP-1 and U937 for 48 h, the protein levels of Arginase-1 and CD163 were determined through Western blotting. *P < 0.05, **P < 0.01, ***P < 0.001 vs. OE-NC. C The ratio of CD206-positive or CD163-positive cells following transfection of OE-SNORD52/ OE-NC was analyzed by flow cytometry. ***P < 0.001 vs. OE-NC. D Following transfec- tion of OE-SNORD52/OE-NC in THP-1 and U937 for 48 h, the levels of JAK2/STAT6 pathway-related proteins were determined through Western blotting. ***P < 0.001 vs. OE-NC. E The IL-4 and IL-13 treated THP-1/U937 mac- rophages were co-incubated with SNORD52 ASO-1/-2 or ASO-NC for 48 h, and then the mRNA expression of SNORD52 was detected by qRT-PCR. *P < 0.05, **P < 0.01, ***P < 0.001 vs. ASO-NC. F The IL-4 and IL-13 treated THP-1/U937 mac- rophages were co-incubated with SNORD52 ASO-1/-2 or ASO-NC for 48 h, and then the protein levels of Arginase-1 and CD163 were determined through Western blotting. ***P < 0.001 vs. ASO-NC. G The ratio of CD206-positive or CD163-positive cells following transfection of SNORD52- ASO-1/-2 or ASO-NC was analyzed by flow cytometry. ***P < 0.001 vs. ASO-NC. (H) The IL-4 and IL-13 treated THP-1/ U937 macrophages were co-incubated with SNORD52 ASO-1/-2 or ASO-NC for 48 h, and then the levels of JAK2/STAT6 pathway-related proteins were determined through Western blotting. ***P < 0.001 vs. ASO-NC. ns: no significance

Article Snippet: Meanwhile, the primary antibodies Arginase-1, CD163, and STAT6 used in Western blotting were also purchased from Proteintech, while other primary antibodies including JAK2, p-JAK2, p-STAT6, exosome-positive markers (CD81 and TSG101), and the corresponding HRP-conjugated secondary antibodies were available from Abcam (Cambridge, UK).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Incubation

Journal: Discover oncology

Article Title: Hepatoma cell-derived exosomal SNORD52 mediates M2 macrophage polarization by activating the JAK2/STAT6 pathway.

doi: 10.1007/s12672-024-01700-y

Figure Lengend Snippet: Fig. 5 Huh7 cells-derived exosomal SNORD52 mediates M2 macrophage polarization through activating JAK2/STAT6 pathway. A Following the establishment of co-culture model, AG-490 (a specific JAK2 inhibitor) was added and the levels of JAK2/STAT6 pathway-related pro- teins in receptor THP-1 macrophages were measured by Western blotting. B Following the establishment of co-culture model, AG-490 (a specific JAK2 inhibitor) was added and the ratio of CD206-positive or CD163-positive THP-1 macrophages was analyzed by flow cytometry. **P < 0.01, ***P < 0.001. ns: no significance

Article Snippet: Meanwhile, the primary antibodies Arginase-1, CD163, and STAT6 used in Western blotting were also purchased from Proteintech, while other primary antibodies including JAK2, p-JAK2, p-STAT6, exosome-positive markers (CD81 and TSG101), and the corresponding HRP-conjugated secondary antibodies were available from Abcam (Cambridge, UK).

Techniques: Derivative Assay, Co-Culture Assay, Western Blot, Flow Cytometry

Journal: Pathology and Oncology Research

Article Title: Pathological Changes and Expression of JAK-STAT Signaling Pathway Hallmark Proteins in Rat Retinas at Different Time Points After Retinal Ischemia Reperfusion Injury

doi: 10.3389/pore.2022.1610385

Figure Lengend Snippet: Immunohistochemistry staining showing protein expression and localization of JAK2, STAT3, p-JAK2, p-STAT3, Bcl-2, and Bax in control, RIRI 0, 6, 24, 72, and 144 h groups. (A) Protein expression of JAK2, STAT3, p-JAK2, p-STAT3, Bcl-2, and Bax in six groups. Scale bar = 50 μm. (B–G) Analysis of JAK2 (B) , STAT3 (C) , p-JAK2 (D) , p-STAT3 (E) , Bcl-2 (F) , and Bax (G) in six groups. N = 5 sections for measuring per group. Compared with control group; ns, no significance, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies were as follows: JAK2 (bs-23003r, Bioss), STAT3 (60199-1-lg, Proteintech), p-JAK2 (ab32101, Abcam), p-STAT3 (sc-8059, Santa), Bcl-2 (ab194583, Abcam), and Bax (A19684, ABclonal).

Techniques: Immunohistochemistry, Staining, Expressing

Journal: Pathology and Oncology Research

Article Title: Pathological Changes and Expression of JAK-STAT Signaling Pathway Hallmark Proteins in Rat Retinas at Different Time Points After Retinal Ischemia Reperfusion Injury

doi: 10.3389/pore.2022.1610385

Figure Lengend Snippet: The hallmarker protein expression of JAK-STAT signaling pathway in control, RIRI 0, 6, 24, 72, and 144 h groups. (A) Protein expression of JAK2, p-JAK2, STAT3, and p-STAT3 in six time points groups. (B–E) Analysis of JAK2 (B) , p-JAK2 (C) , STAT3 (D) , and p-STAT3 (E) in six groups. N = 3–6 repeats per group. Compared with control group, ns, no significance, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies were as follows: JAK2 (bs-23003r, Bioss), STAT3 (60199-1-lg, Proteintech), p-JAK2 (ab32101, Abcam), p-STAT3 (sc-8059, Santa), Bcl-2 (ab194583, Abcam), and Bax (A19684, ABclonal).

Techniques: Expressing