control shrna  (Santa Cruz Biotechnology)

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

Journal: Acta Neuropathologica

Article Title: TDP-43 impairs glycolysis by sequestering hexokinase 1 in amyotrophic lateral sclerosis

doi: 10.1007/s00401-026-02996-6

Figure Lengend Snippet: Pathological TDP-43 elicits glycolytic impairment in cells. a , b Agilent seahorse glycolytic stress test was performed on n = 3 independent biological repeats. Glycolysis and Glycolytic Capacity were measured according to manufacturer’s protocol. ECAR values were normalized to total protein then plotted against time. Individual glycolysis and glycolytic capacity values were normalized to total protein level then to individual control from each biological repeat. a Stable HEK293 cells were overexpressed with either YFP (control), TDP-43 WT or TDP-43 ΔNLS . Data were analyzed by one-way ANOVA followed by post hoc Tukey’s test. b TDP-43 G298S patient iPSC and control iPSC were differentiated into motor neurons with our established protocol. On day 26 following differentiation, cells were treated with MG-132 (1 μM) for 24 h. Data were analyzed by Student’s t-test. c YFP (control), TDP-43 WT or TDP-43 ΔNLS stable cells were transiently transfected with various glycolysis shRNAs. Cell viability was measured via MTT assay. Decreased cell viability in all TDP-43 WT and TDP-43 ΔNLS with various glycolysis shRNA (n = 3). Data were analyzed by one-way ANOVA followed by post hoc Tukey’s test. All data are mean ± SE

Article Snippet: The following constructs pcDNA3.2-YFP (addgene #84910), pcDNA3.2-TDP-43 WT -YFP (human) (addgene #84911), pcDNA3.2-TDP-43 ΔNLS -YFP (human) (addgene #84912), pLD-puro-Cc-TARDBP-A315T_VA Plasmid (human TDP-43 A315T) (addgene #141329), pLD-puro-Cc-TARDBP-WT_VA Plasmid (human TDP-43 WT) (addgene #141327), and shRNA control (addgene: #8453) were purchased from addgene.

Techniques: Control, Transfection, MTT Assay, shRNA

Journal: Cell Death Discovery

Article Title: SPINK2 silencing suppresses leukemic proliferation and restores myeloid commitment via MECOM downregulation in acute myeloid leukaemia

doi: 10.1038/s41420-026-02988-1

Figure Lengend Snippet: A Barplot showing the expression of SPINK2 measured by qPCR in a panel of AML cell lines. n = 6 for each cell line. B Bowtie plot representing SPINK2 mRNA quantification by q-PCR following SPINK2 shRNA silencing and normalized using GAPDH ( n = 6). C Lineplot displaying cell viability upon SPINK2 knock-down determined by counting cells every 24 h for four consecutive days post shRNA induction. The results are indicative of 3 independent experiments ( n = 6). D Two-dimensional flow cytometric dot plot showing BrdU incorporation 72 hours post doxycycline induction. The three gates indicate the G0/G1, S and G2/M phases of the cell cycle. The barplot on the righthand side represent an average of three independent experiments ( n = 6). E Bowtie plot showing the percentages of Annexin V + apoptotic/necrotic cells. F Representative two-dimensional flow cytometric contour plots showing the expression levels of CD14, CD64, CD35 and CD300E in FUJIOKA cells that have undergone SPINK2 silencing 96 hours post doxycycline induction ( n = 6). The statistical analysis reported in this figure have been performed using student’s t test (*** p < 0.001, * p < 0.05).

Article Snippet: A scrambled control shRNA plasmid (shCtr), Tet-pLKO-puro-Scrambled (Addgene plasmid #47541), was used as a non-targeting control [ ].

Techniques: Expressing, shRNA, Knockdown, BrdU Incorporation Assay