Journal: bioRxiv

Article Title: Ruxolitinib clears CRYAB p.Arg120Gly aggregates through the ubiquitin-proteasome system

doi: 10.1101/2024.10.11.615348

Figure Lengend Snippet: NRVMs were transduced with AdV5-CMV-CRYAB R120G -GFP and treated with 3 µM ruxolitinib or DMSO or 100 nM siRNAs targeting either JAK1 (siJak1), JAK2 (siJak2), STAT3 (siStat3) or scramble siRNA (scr). Thereafter, NRVMs were harvested or fixed after 4-6 days. A , Treatment with ruxolitinib or DMSO; Scale bar = 100 µm. B , Transfection with siJak1; Scale bar = 200 µm. C , Transfection with siJak2; Scale bar = 100 µm D , Transfection with siStat3, Scale bar = 20 µm. A , B , C , D , Western blots of protein extracts from treated NRVMs were stained with antibodies directed against indicated proteins. In the representative immunofluorescence images, aggregates are depicted in magenta (CRYAB R120G -GFP), cardiomyocytes in yellow (anti-cardiac troponin I), and nuclei in blue (DAPI). Quantification of aggregates in cardiomyocytes with NIS Elements or ImageJ software. Data were obtained from 1 (Western blot) or 2 (immunofluorescence) NRVM preparations with at least 3 wells per condition and at least 7 images per well for immunofluorescence. Data are depicted as mean ± SEM, and p-values were obtained with the unpaired Student’s t-test. Abbreviation: ns, non-significant.

Article Snippet: RT-qPCR was then performed with TaqMan gene expression assays (Thermo Fisher Scientific; Psmd1 - Rn01400483_m1, Jak1 - Mm00600614_m1, Jak2 -Mm01208489_m1, Stat1 - Mm01257286_m1, Stat3 - Mm01219775_m1, Tyk2 -Mm00444469_m1, 18s - Hs03003631_g1) and SsoAdvancedTM Universal Probes Supermix (BioRad #1725281).

Techniques: Transduction, Transfection, Western Blot, Staining, Immunofluorescence, Software

Journal: bioRxiv

Article Title: Ruxolitinib clears CRYAB p.Arg120Gly aggregates through the ubiquitin-proteasome system

doi: 10.1101/2024.10.11.615348

Figure Lengend Snippet: A , Heart weight-to-body weight ratio (HW/BW) in 1-, 4- and 7-month-old CRYAB p.Arg120Gly transgenic (R120G) and non-transgenic (NTG) mice. B , Ejection fraction (EF) in 7-month-old R120G and NTG mice. C , Representative Western blots of phosphorylated (P-)STAT3, STAT3 and indicated control in 1-, 4- and 7-month-old R120G and NTG mice. D , P-STAT3/STAT3 quantification of Western blots from 1-, 4- and 7-month-old R120G and NTG mice. E , Stat3 mRNA level determined by RT-qPCR from 7-month-old R120G and NTG mice. Western blot quantification was performed with Image Lab software. Data are depicted as mean ± SEM, and p-values were obtained with the unpaired Student’s t-test. Abbreviation: ns, non-significant.

Article Snippet: RT-qPCR was then performed with TaqMan gene expression assays (Thermo Fisher Scientific; Psmd1 - Rn01400483_m1, Jak1 - Mm00600614_m1, Jak2 -Mm01208489_m1, Stat1 - Mm01257286_m1, Stat3 - Mm01219775_m1, Tyk2 -Mm00444469_m1, 18s - Hs03003631_g1) and SsoAdvancedTM Universal Probes Supermix (BioRad #1725281).

Techniques: Transgenic Assay, Western Blot, Control, Quantitative RT-PCR, Software

Journal: bioRxiv

Article Title: Ruxolitinib clears CRYAB p.Arg120Gly aggregates through the ubiquitin-proteasome system

doi: 10.1101/2024.10.11.615348

Figure Lengend Snippet: CRYAB p.Arg120Gly transgenic (R120G) and non-transgenic (NTG) mice treated for 3 weeks with 75 mg/kg ruxolitinib (ruxo) or vehicle (veh) twice-daily oral gavage. Transthoracic echocardiography was performed at the start (before, 21-week-old) and the end of the treatment (veh or ruxo, 24-week-old). A , Scheme of experimental outline. B , WB and quantification of phosphorylated (P-) STAT3, STAT3 and indicated controls at the end of treatment. WB quantification was performed with Image Lab software. C, Ejection fraction (EF) and left ventricular mass-to-body weight ratio (LVM/BW) before and after (veh/ruxo) treatment. D , Heart weight-to-body weight ratio (HW/BW) and body weight (BW) at the end of the treatment. E , Representative images and quantification of R120G and NTG mouse heart sections after vehicle or ruxolitinib treatment. CRYAB is depicted in green, cardiomyocytes in purple (anti-cardiac troponin I) and nuclei in blue (DAPI). Scale bar = 10 µm. Quantification of aggregates in cardiomyocytes with NIS Elements software. At least 5 images of 3 mice per group were analyzed. Data are depicted as mean ± SEM, and p-values were obtained with the one- way ( B ) or two-way ANOVA and Tukey’s multiple comparisons post-hoc analysis ( C, D ) or unpaired Student’s t-test ( E ). Abbreviation: ns, non-significant.

Article Snippet: RT-qPCR was then performed with TaqMan gene expression assays (Thermo Fisher Scientific; Psmd1 - Rn01400483_m1, Jak1 - Mm00600614_m1, Jak2 -Mm01208489_m1, Stat1 - Mm01257286_m1, Stat3 - Mm01219775_m1, Tyk2 -Mm00444469_m1, 18s - Hs03003631_g1) and SsoAdvancedTM Universal Probes Supermix (BioRad #1725281).

Techniques: Transgenic Assay, Software

Journal: bioRxiv

Article Title: Ruxolitinib clears CRYAB p.Arg120Gly aggregates through the ubiquitin-proteasome system

doi: 10.1101/2024.10.11.615348

Figure Lengend Snippet: Heterozygous (het) or homozygous (hom) Jak1 knockout (KO) was induced in MCM-transgenic (TG) mice crossed with CRYAB p.Arg120Gly TG (R120G) or non-transgenic (NTG) mice with tamoxifen chow. Transthoracic echocardiography was performed 26 weeks and hearts were extracted. A , Scheme of experimental outline. B , Jak1 mRNA levels determined by RT-qPCR. C , JAK1 protein levels determined by Western blot and normalized to ACTN2. Western blot quantification was performed with Image Lab software D, Ejection fraction (EF) and left ventricular mass-to-body weight ratio (LVM/BW). E , Heart weight-to-body weight ratio (HW/BW) and body weight (BW). F , Representative images and quantification of mouse heart sections. Aggregates are depicted in green, cardiomyocytes in purple (anti-cardiac troponin I) and nuclei in blue (DAPI). Scale bar = 50 µm. Quantification of aggregates in cardiomyocytes with NIS Elements software. At least 5 images of 3 mice per group were analyzed. Data are depicted as mean ± SEM, and p-values were obtained with the one-way ( B ) or two- way ANOVA ( D , E ) with Tukey’s multiple comparisons post-hoc analysis or unpaired Student’s t-test ( C , F ). Abbreviation: ns, non-significant.

Article Snippet: RT-qPCR was then performed with TaqMan gene expression assays (Thermo Fisher Scientific; Psmd1 - Rn01400483_m1, Jak1 - Mm00600614_m1, Jak2 -Mm01208489_m1, Stat1 - Mm01257286_m1, Stat3 - Mm01219775_m1, Tyk2 -Mm00444469_m1, 18s - Hs03003631_g1) and SsoAdvancedTM Universal Probes Supermix (BioRad #1725281).

Techniques: Knock-Out, Transgenic Assay, Quantitative RT-PCR, Western Blot, Software

Journal: Cancer medicine

Article Title: Direct effect of dasatinib on signal transduction pathways associated with a rapid mobilization of cytotoxic lymphocytes.

doi: 10.1002/cam4.925

Figure Lengend Snippet: Figure 1. Flow cytometric analysis of each lymphocyte subset and expression of phosphorylated proteins. Representative data are shown. Lymphocyte fractions are classified according to surface antibodies including CD3, CD8, and CD56. Natural killer (NK) cells were defined as the CD3-CD56+ immunophenotype and cytotoxic T lymphocytes (CTLs) were CD3+ CD8+ (A). Cells were costained with phospho-specific antibodies, including antibodies targeting pJAK1, pJAK2, pSTAT1, pSTAT3, pERK, pJNK, pp38, and pAKT, and expression levels of the isotype control and phosphorylated proteins in NK cells are presented (B). The values for the isotype control and each phosphorylated protein are shown as the median fluorescence intensity (MFI).

Article Snippet: Fluorescein isothiocyanate (FITC)- labeled polyclonal rabbit IgG antibodies for phosphorylated proteins, including pJAK1 (Y1034, #3238R), pJAK2 (Y1007, #2485R), pSTAT1 (Y701, #1657R), pSTAT3 (Y705, #1658R), pERK (T202/Y204, #1646R), pJNK (T183/Y185, #1640R), pp38 (T180/Y182, #2210R), pAKT (Y315, #5193R), and isotype control (#0295P) were purchased from Bioss (Wobun, MA, USA).

Techniques: Expressing, Control, Fluorescence

Journal: Cancer medicine

Article Title: Direct effect of dasatinib on signal transduction pathways associated with a rapid mobilization of cytotoxic lymphocytes.

doi: 10.1002/cam4.925

Figure Lengend Snippet: Figure 3. Constitutive levels of phosphorylated proteins including pJAK1, pJAK2, pSTAT1, pSTAT3, pERK, pJNK, pp38, and pAKT in natural killer (NK) cells (A) and cytotoxic T lymphocytes (CTLs) (B) grouped according to treatment (dasatinib [n = 18] or other TKI [n = 12]) are shown. The values for phosphorylated proteins in each fraction are shown as the median fluorescence intensity (MFI).

Article Snippet: Fluorescein isothiocyanate (FITC)- labeled polyclonal rabbit IgG antibodies for phosphorylated proteins, including pJAK1 (Y1034, #3238R), pJAK2 (Y1007, #2485R), pSTAT1 (Y701, #1657R), pSTAT3 (Y705, #1658R), pERK (T202/Y204, #1646R), pJNK (T183/Y185, #1640R), pp38 (T180/Y182, #2210R), pAKT (Y315, #5193R), and isotype control (#0295P) were purchased from Bioss (Wobun, MA, USA).

Techniques: Fluorescence

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: P orphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by activating the neutrophil chemotaxis in the tumour microenvironment

doi: 10.1007/s00262-022-03348-5

Figure Lengend Snippet: Acquisition of the EMT phenotype was verified by activation of the JAK1/STAT3 signalling pathway through the CXCL2/CXCR2 signalling axis. a Western blot analysis showed that EMT occurred and the JAK1/STAT3 signalling pathway was activated by CXCL2/CXCR2 when P. gingivalis infected the TME of OSCC. b After coculture for 24 h, and the cells were transinfected with sh-CXCL2 and sh-CXCR2 lentiviruses for 72 h, the cells were observed under a fluorescence microscope. c, d Knockdown of CXCL2 and CXCR2 was verified at the protein and mRNA levels by Western blotting and qRT-PCR

Article Snippet: Primary antibodies against P. gingivalis (Dia-An, lnc, it was freshly prepared, 1:1000), CXCL2 (bs-1162R, BIOSS, Biotech, Beijing, China, 1:2000), CXCR2 (bs-1629R, BIOSS, Biotech, Beijing, China, 1:2000), JAK1 (bs-1439R, BIOSS, Biotech, Beijing, China, 1:2000), STAT3 (bs-55208R, BIOSS, Biotech, Beijing, China, 1:2000), p-JAK1 (bs-3238R, BIOSS, Biotech, Beijing, China, 1:2000), p-STAT3 (bs-1658R, BIOSS, Biotech, Beijing, China, 1:2000), E-cadherin (bs-10009R, BIOSS, Biotech, Beijing, China, 1:2000), N-cadherin (bs-1172R, BIOSS, Biotech, Beijing, China, 1:2000), and GAPDH (bs-13282R, BIOSS, Biotech, Beijing, China, 1:2000) and secondary antibodies (anti‐rabbit‐HRP, Cell Signalling Technology, Inc, Boston, USA) were used in this study.

Techniques: Activation Assay, Western Blot, Infection, Fluorescence, Microscopy, Knockdown, Quantitative RT-PCR

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: P orphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by activating the neutrophil chemotaxis in the tumour microenvironment

doi: 10.1007/s00262-022-03348-5

Figure Lengend Snippet: Screening and culture of positive clone cells expressing both the lentiviruses of sh-CXCL2 and sh-CXCR2 which can reverse the phenotype of EMT and rescue the signalling pathway of JAK1/STAT3. a The lentiviruses of sh-CXCL2 and sh-CXCR2 were co-transfected into TSCCa cells. b Flow cytometry was used to screen out the positive cells that expression both lentiviruses of sh-CXCL2 and sh-CXCR2. c The clone of positive cells were screened out and cultured by limited dilution method. d Western blot analysis after inactivation of the CXCL2/CXCR2 signalling axis in the P. gingivalis-infected TME of OSCC, the activity of the JAK1/STAT3 signalling pathway was decreased, and the EMT process was reversed. (*p < 0.05, **p < 0.01, and ***p < 0.001; one‐way analysis of variance)

Article Snippet: Primary antibodies against P. gingivalis (Dia-An, lnc, it was freshly prepared, 1:1000), CXCL2 (bs-1162R, BIOSS, Biotech, Beijing, China, 1:2000), CXCR2 (bs-1629R, BIOSS, Biotech, Beijing, China, 1:2000), JAK1 (bs-1439R, BIOSS, Biotech, Beijing, China, 1:2000), STAT3 (bs-55208R, BIOSS, Biotech, Beijing, China, 1:2000), p-JAK1 (bs-3238R, BIOSS, Biotech, Beijing, China, 1:2000), p-STAT3 (bs-1658R, BIOSS, Biotech, Beijing, China, 1:2000), E-cadherin (bs-10009R, BIOSS, Biotech, Beijing, China, 1:2000), N-cadherin (bs-1172R, BIOSS, Biotech, Beijing, China, 1:2000), and GAPDH (bs-13282R, BIOSS, Biotech, Beijing, China, 1:2000) and secondary antibodies (anti‐rabbit‐HRP, Cell Signalling Technology, Inc, Boston, USA) were used in this study.

Techniques: Expressing, Transfection, Flow Cytometry, Cell Culture, Western Blot, Infection, Activity Assay

Journal: Cell Death & Disease

Article Title: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

doi: 10.1038/s41419-021-04021-y

Figure Lengend Snippet: a qRT-PCR analysis of miR-4293 levels in NSCLC tissues and corresponding para-carcinoma tissues ( n = 16). Data are expressed as median (interquartile range). ** P < 0.01; Mann–Whitney U test. b qRT-PCR analysis of miR-4293 levels in A549, H1299, H1975, and HBE cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. c qRT-PCR analysis of miR-4293 level in A549 cells with transfection of miR-4293 mimic, ASO-4293, mu-4293 (mutant), or scrambled. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. d MTT assay of A549 at 24 h post-transfection of miR-4293, ASO-4293, mu-4293 or scrambled. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01, * p < 0.05; ANOVA test. e FACS analysis of A549 cell apoptosis at 24 h post-transfection of miR-4293, ASO-4293, mu-4293, or scrambled. f Western blot detection of STAT3 and apoptosis-related factors (Bcl-2, Bax, BIRC3, and Caspase 3,8,9) in miR-4293-, ASO-4293-, or scramble-treated A549 cells. g , h Transwell migration assay of A549 at 24 h post-transfection of miR-4293, ASO-4293, mu-4293, or scrambled. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. i , j Colony formation assays of A549 at 24 h post-transfection of miR-4293, ASO-4293, mu-4293, or scrambled. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test.

Article Snippet: The antibodies used were as follows: rabbit anti-human DCP2 (1:1000; ab28658; abcam, Ltd), rabbit anti-human STAT3 (1:500; bsm-33218M; Bioss, Ltd), rabbit antihuman p-STAT3 (1:800; AP0248), mouse anti-human BIRC3 (1:500; MB0129), rabbit anti-human Bax (1:500; BS6420), Bcl2 (1:500; BS70205), rabbit anti-human Caspase 3 (1:1000; BS61583), rabbit anti-human Caspase 8(1:1000; AP0258), rabbit anti-human Caspase 9 (1:1000; AP0359), and rabbit anti-human GAPDH (1:3000; MB001) all from Bioworld Technology, Ltd).

Techniques: Quantitative RT-PCR, MANN-WHITNEY, Transfection, Mutagenesis, MTT Assay, Western Blot, Transwell Migration Assay

Journal: Cell Death & Disease

Article Title: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

doi: 10.1038/s41419-021-04021-y

Figure Lengend Snippet: a TargetScan analysis predicted that the DCP2 mRNA-3′-UTR is targeted by miR-4293. We synthesized a mutant miR-4293 (Mu-4293) and cloned DCP2 mRNA-3’-UTR and mu-DCP2–3’-UTR sequences. b Analysis of miR-4293 regulating luciferase expression of reporter plasmids containing DC2–3’UTR (right panel) or mu-DCP2–3’UTR (left panel). Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. c Analysis of ASO-4293 regulating luciferase expression of reporter plasmids containing DC2–3’UTR (right panel) or mu-DCP2–3’UTR (left panel). Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. d Western blot detection of DCP2 in miR-4293-, ASO-4293-, or scramble-treated A549 cells. e Western blot detection of DCP2 in siRNA-DCP2–1-, siRNA-DCP2–2-, or scramble-treated A549 cells and H129 cells. f qRT-PCR analysis of WFDC21P level in pcDNA-WFDC21P + scramble-, si-DCP2-, si-DCP2 + pcDNA-WFDC21P-, or scramble-treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. g Western blot detection of STAT3 phosphorylation in pcDNA-WFDC21P + scramble-, si-DCP2-, si-DCP2 + pcDNA-WFDC21P-, or scramble-treated A549 cells. h qRT-PCR analysis of DCP2 level in pcDNA or pcDNA-DCP2 -treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. i qRT-PCR analysis of WFDC21P level in 0 h, 4 h, 8 h, 12 h, 24 h, 48 h after transfection of pcDNA-DCP2. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. j RIP assay of the interaction between WFDC21P and DCP2. Upper panel, qRT-PCR analysis of WFDC21P pull-down by IgG or DCP2; lower panel, electrophoresis for WFDC21P pull-down by IgG or DCP2 after PCR amplification. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. k qRT-PCR analysis of WFDC21P levels in miR-4293-, ASO-4293, Mu-4293 or scramble-treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. l qRT-PCR analysis of WFDC21P levels in miR-4293+pcDNA-WFDC21P-, or scramble+ pcDNA-WFDC21P-treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. m Western blot detection of STAT3 phosphorylation in pcDNA-WFDC21P + scramble-, miR-4293-, miR-4293+pcDNA-WFDC21P-, or scramble-treated A549 cells. n Western blot detection of STAT3 phosphorylation in AG490 + pcDNA-, pcDNA-WFDC21P-, AG490 + pcDNA-WFDC21P-, or pcDNA treated A549 cells. o qRT-PCR analysis of miR-4293 level in AG490 + pcDNA-, pcDNA-WFDC21P-, AG490 + pcDNA-WFDC21P-, or pcDNA treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01, * P < 0.05; ANOVA test.

Article Snippet: The antibodies used were as follows: rabbit anti-human DCP2 (1:1000; ab28658; abcam, Ltd), rabbit anti-human STAT3 (1:500; bsm-33218M; Bioss, Ltd), rabbit antihuman p-STAT3 (1:800; AP0248), mouse anti-human BIRC3 (1:500; MB0129), rabbit anti-human Bax (1:500; BS6420), Bcl2 (1:500; BS70205), rabbit anti-human Caspase 3 (1:1000; BS61583), rabbit anti-human Caspase 8(1:1000; AP0258), rabbit anti-human Caspase 9 (1:1000; AP0359), and rabbit anti-human GAPDH (1:3000; MB001) all from Bioworld Technology, Ltd).

Techniques: Synthesized, Mutagenesis, Clone Assay, Luciferase, Expressing, Western Blot, Quantitative RT-PCR, Transfection, Electrophoresis, Amplification

Journal: Cell Death & Disease

Article Title: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

doi: 10.1038/s41419-021-04021-y

Figure Lengend Snippet: a qRT-PCR analysis for WFDC21P level in NSCLC tissues and corresponding para-carcinoma tissues ( n = 16). Data are expressed as median (interquartile range). ** P < 0.01; Mann–Whitney U test. b qRT-PCR analysis for WFDC21P level in A549, H1299, H1975, and HBE cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. c Western blot detection of STAT3 in A549, H1299, H1975, and HBE cells. d qRT-PCR analysis of STAT3 expression in NSCLC tissues and corresponding para-carcinoma tissues ( n = 16). Data are expressed as median (interquartile range). ** P < 0.01; Mann–Whitney U test. e – g Pearson′s correlation analysis of miR-4293 expression and WFDC21P expression ( n = 16, ** P = 0.015) ( e ), miR-4293 expression and STAT3 expression (n = 16, ** P < 0.01) ( f ), WFDC21P expression and STAT3 expression ( n = 16, ** P < 0.01) ( g ).

Article Snippet: The antibodies used were as follows: rabbit anti-human DCP2 (1:1000; ab28658; abcam, Ltd), rabbit anti-human STAT3 (1:500; bsm-33218M; Bioss, Ltd), rabbit antihuman p-STAT3 (1:800; AP0248), mouse anti-human BIRC3 (1:500; MB0129), rabbit anti-human Bax (1:500; BS6420), Bcl2 (1:500; BS70205), rabbit anti-human Caspase 3 (1:1000; BS61583), rabbit anti-human Caspase 8(1:1000; AP0258), rabbit anti-human Caspase 9 (1:1000; AP0359), and rabbit anti-human GAPDH (1:3000; MB001) all from Bioworld Technology, Ltd).

Techniques: Quantitative RT-PCR, MANN-WHITNEY, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

doi: 10.1038/s41419-021-04021-y

Figure Lengend Snippet: a MTT assays of A549 at 24 h post-transfection of pcDNA-WFDC21P and pc-DNA. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. b qRT-PCR analysis of WFDC21P level in si-WFDC21P- and scramble-treated cells. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. c MTT assays of A549 at 24 h post-transfection of si-WFDC21P and scramble. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. d FACS analysis of A549 cell apoptosis at 24 h post-transfection of pc-DNA, pcDNA-WFDC21P, si-WFDC21P, and scrambled control. e Western blot detection of STAT3 and apoptosis-related factors (Bcl-2, Bax, BIRC3, and Caspase 3,8,9) pcDNA vs pcDNA-WFDC21P; scramble vs si-WFDC. f – i Transwell assays of A549 migrat i on. f , g pcDNA vs pcDNA-WFDC21P; ( h ) and ( i ) scramble vs. si-WFDC. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test. j – m Colony formation assays of A549. j , k pcDNA vs pcDNA-WFDC21P; ( l ) and ( m ) scramble vs. si-WFDC. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; Student’s t -test.

Article Snippet: The antibodies used were as follows: rabbit anti-human DCP2 (1:1000; ab28658; abcam, Ltd), rabbit anti-human STAT3 (1:500; bsm-33218M; Bioss, Ltd), rabbit antihuman p-STAT3 (1:800; AP0248), mouse anti-human BIRC3 (1:500; MB0129), rabbit anti-human Bax (1:500; BS6420), Bcl2 (1:500; BS70205), rabbit anti-human Caspase 3 (1:1000; BS61583), rabbit anti-human Caspase 8(1:1000; AP0258), rabbit anti-human Caspase 9 (1:1000; AP0359), and rabbit anti-human GAPDH (1:3000; MB001) all from Bioworld Technology, Ltd).

Techniques: Transfection, Quantitative RT-PCR, Western Blot

Journal: Cell Death & Disease

Article Title: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

doi: 10.1038/s41419-021-04021-y

Figure Lengend Snippet: The above-mentioned data showed that miR-4293 transfection elevated miR-4293 levels, which could be attenuated by ASO. pcDNA-WFDC21P transfection increased WFDC21P. Si-WFDC treatment reduced WFDC21P level. a MTT assay of A549 at 24 h post-transfection of si-WFDC + scramble, miR-4293, miR-4293+si-WFDC, and scramble. Data are expressed as mean ± SD for triplicate experiments. ** p < 0.01; ANOVA test. b FACS analysis of A549 cell apoptosis at 24 h post-transfection of si-WFDC + scramble, miR-4293, miR-4293+si-WFDC, and scramble. c Western blot detection p-STAT3, Bcl-2, BAX, cleaved-caspase 3, 8, and 9 in si-WFDC + scramble-, miR-4293-, miR-4293+si-WFDC- or scramble-treated cells. d , e RTCA station analysis of migration for A549 with transcription of si-WFDC + scramble-, miR-4293-, miR-4293+si-WFDC- or scramble. f MTT of A549 at 24 h post-transfection of pcDNA+ASO-4293, pcDNA-WFDC21P + scramble, pcDNA-WFDC21P + ASO-4293, and pcDNA+ scramble. Data are expressed as mean ± SD for triplicate experiments. (* P < 0.05). g FACS analysis of A549 cell apoptosis at 24 h post-transfection of pcDNA+ASO-4293, pcDNA-WFDC21P + scramble, pcDNA-WFDC21P + ASO-4293, and pcDNA+ scramble. h Western blot detection of STAT3 phosphorylation and apoptosis-related factors (Bcl-2, Bax and Caspase 3) in pcDNA+ASO-4293, pcDNA-WFDC21P + scramble, pcDNA-WFDC21P + ASO-4293, and pcDNA+ scramble-treated A549 cells. i – l RTCA station analysis of migration for A549. i , j pcDNA-WFDC21P vs pc-DNA; ( k ) and ( l ), pcDNA-WFDC21P + scramble vs. pcDNA-WFDC21P + ASO-4293. Data are expressed as mean ± SD for triplicate experiments. * p < 0.05, *** p < 0.001; Student’s t -test.

Article Snippet: The antibodies used were as follows: rabbit anti-human DCP2 (1:1000; ab28658; abcam, Ltd), rabbit anti-human STAT3 (1:500; bsm-33218M; Bioss, Ltd), rabbit antihuman p-STAT3 (1:800; AP0248), mouse anti-human BIRC3 (1:500; MB0129), rabbit anti-human Bax (1:500; BS6420), Bcl2 (1:500; BS70205), rabbit anti-human Caspase 3 (1:1000; BS61583), rabbit anti-human Caspase 8(1:1000; AP0258), rabbit anti-human Caspase 9 (1:1000; AP0359), and rabbit anti-human GAPDH (1:3000; MB001) all from Bioworld Technology, Ltd).

Techniques: Transfection, MTT Assay, Western Blot, Migration

Journal: Cell Death & Disease

Article Title: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

doi: 10.1038/s41419-021-04021-y

Figure Lengend Snippet: miR-4293 promotes tumor cell proliferation and metastasis but suppresses apoptosis. DCP2 can directly or indirectly bind to and downregulate WFDC21P. By targeting DCP2 mRNA, miR-4293 suppresses the expression of DCP2, and further enhances the expression of WFDC21P and promotes STAT3 phosphorylation. As a result of elevated STAT3 activity, apoptosis is suppressed and proliferation is enhanced through Bcl2 elevation, Bax2 attenuation and reduced caspase cleavage. In addition, WFDC21P elevates miR-4293 expression by promoting STAT3 activation.

Article Snippet: The antibodies used were as follows: rabbit anti-human DCP2 (1:1000; ab28658; abcam, Ltd), rabbit anti-human STAT3 (1:500; bsm-33218M; Bioss, Ltd), rabbit antihuman p-STAT3 (1:800; AP0248), mouse anti-human BIRC3 (1:500; MB0129), rabbit anti-human Bax (1:500; BS6420), Bcl2 (1:500; BS70205), rabbit anti-human Caspase 3 (1:1000; BS61583), rabbit anti-human Caspase 8(1:1000; AP0258), rabbit anti-human Caspase 9 (1:1000; AP0359), and rabbit anti-human GAPDH (1:3000; MB001) all from Bioworld Technology, Ltd).

Techniques: Expressing, Activity Assay, Activation Assay

Journal: Oncotarget

Article Title: Structural composition of components of geoherb Moutan Cortex contributes to anti-diabetic nephropathy activity

doi: 10.18632/oncotarget.23771

Figure Lengend Snippet: Figure 1: Comparison on the down-regulation of Moutan Cortex from different regions on protein expression levels of ICAM-1,TGF-β1 and FN. HBZY-1 mesangial cells were treated with 200 μg/mL AGEs in the presence or absence of Moutan Cortex (MC) extract of 200 μg/mL. Aminoguanidine of 10 μM was used as the positive control while BSA (200 μg/mL) as blank control. (A) Western blotting was performed to compare the protein expression levels. (B–D) The grayscale scan results of ICAM-1, TGF-β1 and FN. a, Control; b, 200 μg/mL AGEs; c, Positive control aminoguanidine group; “d-m” represent MC from Anhui, Guizhou, Zhejiang, Henan, Hunan, Hebei, Sichuan, Chongqing, Shandong, Gansu. Data are expressed as means ± SD, n = 3. ###P < 0.001 vs. BSA group; *P < 0.05, **P < 0.01 and ***P < 0.001 vs. AGEs group; $P < 0.05, $$P < 0.01 and $$$P < 0.001 vs. MC from Anhui group.

Article Snippet: Rabbit anti-mouse FN, TGF-β1 and ICAM-1 monoclonal antibodies were offered by Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Comparison, Expressing, Positive Control, Control, Western Blot

Journal: Oncotarget

Article Title: Structural composition of components of geoherb Moutan Cortex contributes to anti-diabetic nephropathy activity

doi: 10.18632/oncotarget.23771

Figure Lengend Snippet: Figure 2: Effect of Moutan Cortex from different regions on ICAM-1 and TGF-β1 protein expression levels in kidney of DN rats. After being treated with STZ and/or MC extract of 5g/kg or , immunohistochemistry was conducted to evaluate the expression levels of ICAM-1 (A) and TGF-β1 (B) of renal tissues. “a” represents normal control; “b” represents model group (DN rats); “c” represents Positive control 0.1 g/kg AG; “d-m” represent Gansu, Chongqing, Shangdong, Sichuan, Zhejiang, Anhui, Hunan, Guizhou, Hebei, Henan.

Article Snippet: Rabbit anti-mouse FN, TGF-β1 and ICAM-1 monoclonal antibodies were offered by Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Control, Positive Control