stat3 sirnas  (Sangon Biotech)

Journal: bioRxiv

Article Title: STING–STAT3–SOX18 Axis Drives EndMT and Epigenetic Reprogramming in SAVI Lung Fibrosis

doi: 10.64898/2026.03.23.713256

Figure Lengend Snippet: A. Top pathways enriched in SAVI P5 vs. P1 (iso-SAVI_SAVI cohort) by Ingenuity Pathway Analysis (IPA). Positive z-scores indicate pathway activation. Additional HC vs. SAVI analysis is shown in Figure S3A–B. B . EndMT signature heatmap in parental SAVI and isogenic iECs, with box-and-whisker plots of representative endothelial, mesenchymal, and previously reported EndMT-associated genes. Asterisks mark genes upregulated under TGFβ-induced EndMT but downregulated in SAVI iECs. C. IPA-derived transcription factor network of significant upstream regulators in the iso-SAVI_SAVI cohort (orange = activated; blue = inhibited). D . Heatmap of activation z-scores for the same transcription factors in panel C, showing similar patterns in SAVI vs. HC iECs and SAVI vs. HC lung tissue. E . Constitutive STAT3 nuclear translocation in SAVI iECs. STAT3 immunofluorescence (red) with DAPI (blue) at P5 from one SAVI and matched isogenic control line; white arrows indicate nuclear STAT3 (scale bar: 20 µm). Quantification reflects STAT3⁺ nuclei per total DAPI⁺ cells across 7–11 fields from 3–4 wells (mean ± SEM; *p < 0.05, two-tailed unpaired t-test). F . STAT3 activation in SAVI lung biopsies. Lung sections from 4 HC and 3 SAVI patients were stained for p-STAT3 Y705 (red) and CD31 (green) with DAPI (blue). Representative images (scale bar: 20 µm) and quantification of p-STAT3 Y705 /DAPI ratios (≥5 images per patient) show increased STAT3 activation in SAVI (mean ± SEM; ***p < 0.001, two-tailed unpaired t-test).

Article Snippet: siRNA plasmids for control (sc-37007), STAT3 (sc-29493), and STAT1 (sc-44123) were purchased from Santa Cruz.

Techniques: Activation Assay, Whisker Assay, Derivative Assay, Translocation Assay, Immunofluorescence, Control, Two Tailed Test, Staining

Journal: bioRxiv

Article Title: STING–STAT3–SOX18 Axis Drives EndMT and Epigenetic Reprogramming in SAVI Lung Fibrosis

doi: 10.64898/2026.03.23.713256

Figure Lengend Snippet: A. Diagram of the Canonical and Non-canonical STING signaling (created with BioRender.com ). B. Constitutive STAT3 and STAT1 activation in SAVI iECs. Phospho-STAT3 Y705 and phospho-STAT1 Y701 (normalized to total protein) were measured in iECs from 3 HC and 3 SAVI donors across passages 1, 3, and 5. SAVI iECs showed increased baseline STAT3 activation (mean ± SEM; ***p < 0.001, **p < 0.01, *p < 0.05, unpaired t-test). C. STING/TBK1 inhibition blocks acute cGAMP-induced STAT3 activation in HC iECs. iECs from 3 HC donors were stimulated with 2’3’-cGAMP (20µg/ml) for 30 min–24 h. STING inhibitor IFM35883 (STINGi, 2.5 μM) and TBK1 inhibitor MRT67307 (TBK1i, 5 μM) were pre-treated one hour before cGAMP. STAT3 activation at 4h was prevented by STING/TBK1 inhibition (mean ±SEM, ***p < 0.001, **p < 0.01, Mann-Whitney test). D. STING/TBK1 inhibition reduces constitutive STAT3 activation in SAVI iECs. SAVI iECs were treated with IFM35883 (2.5 μM) from P2–P5 or with TBK1i (5 μM) for 48 h. Quantification is from four experiments in iEC line from patient SAVI1 (mean ±SEM, ***p < 0.001, unpaired t-test). E. STAT3 shRNA knockdown (KD), not STAT1 KD preserves CD144 (VE-cadherin) in SAVI iECs. iECs from 3 individual SAVI patients infected with control, STAT3, or STAT1 shRNA at P2 were analyzed by flow cytometry at P5 (mean ±SEM, **p < 0.01, paired t-test). Representative flow cytometry profiles are shown in Figure S4C. F. Constitutive protein expression of SLUG/ SNAI2 is elevated in SAVI iECs and suppressed by STING inhibition. Protein from 3 HC, 3 SAVI, and 3 iso-SAVI iECs at P5 showed increased SLUG (normalized by GAPDH) in SAVI; IFM35883 (2.5 μM) treatment from P2–P5 prevented SLUG upregulation in SAVI iECs (SAVI 1) (mean ±SEM, ***p < 0.001, **p < 0.01, unpaired t-test). See also Figure S4E. G. STAT3 shRNA knockdown reduces SLUG mRNA expression (normalized by internal control 18S) in SAVI iECs at P3. Data are summarized from 3 individual SAVI patients iEC lines. *p < 0.01 as determined by two-tailed unpaired t-test.

Article Snippet: siRNA plasmids for control (sc-37007), STAT3 (sc-29493), and STAT1 (sc-44123) were purchased from Santa Cruz.

Techniques: Activation Assay, Inhibition, MANN-WHITNEY, shRNA, Knockdown, Infection, Control, Flow Cytometry, Expressing, Two Tailed Test