control sirna  (Santa Cruz Biotechnology)

Journal: Redox Biology

Article Title: Prophylactic C-terminal occludin–derived peptide attenuates LPS-induced airway inflammation via barrier preservation and mitochondrial ROS regulation

doi: 10.1016/j.redox.2026.104119

Figure Lengend Snippet: The peptide regulates mitochondrial dysfunction and ROS production by inhibiting LPS-induced p38 activation. (A) The BEAS-2b cells were treated with wild-type occludin peptide (pepWT OCLN) or mutant occludin peptide (pepMut OCLN) and incubated with LPS for 15, 30 min. The phospho-specific and total antibodies were assessed by Western blot analysis. β-actin was used as a loading control. (B) The BEAS-2b cells were transfected with p38 overexpression construct (WT p38) or siRNA-p38 for 24 h and incubated with LPS for 4 h siRNA-scramble was used as a negative control. The proinflammatory cytokine transcripts were assessed by qRT-PCR. ∗ p < 0.05 compared with control; ∗∗ p < 0.05 compared with LPS treatment; ∗∗∗ p < 0.05 compared with WT p38-transfected cells. (C) The mitochondrial membrane potential of LPS-induced BEAS-2b cells treated with either WT OCLN peptide or mut peptide was stained with JC-1 dye. Images are representative results of 3 independent experiments. (D) The mitochondria fission was stained using phospho-Drp1 antibody and visualized. The fluorescence intensity was analyzed and statistically evaluated (right panel). ∗ p < 0.05 compared with control; ∗∗ p < 0.05 compared with LPS treatment; ∗∗∗ p < 0.05 compared with WT OCLN peptide-treated transfectants. (E) After the BEAS-2b cells were harvested, cell lysates were used for MTT assay. (F) After mitochondria from the cells was isolated, the mitochondria lysates were used for mtROS measurement. ∗ p < 0.05 compared with control; ∗∗ p < 0.05 compared with LPS only; ∗∗∗ p < 0.05 compared with LPS- and WT occludin peptide-treated cells. All data shown are representative of three independent experiments.

Article Snippet: The siRNA that targets OCLN gene and control siRNA were synthesized by Bioneer (Daejeon, Republic of Korea).

Techniques: Activation Assay, Mutagenesis, Incubation, Western Blot, Control, Transfection, Over Expression, Construct, Negative Control, Quantitative RT-PCR, Membrane, Staining, Fluorescence, MTT Assay, Isolation

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 knockdown suppresses cell proliferation in ESCC cells. (A) Western blot analysis of the protein expression of PRDX4 in KYSE270 cells transfected with PRDX4 siRNA and KYSE30 cells transfected with pcDNA3.1-PRDX4. (B) The relative protein levels of PRDX4 in KYSE270 and KYSE30 cells with different transfections. (C) RT-qPCR assay of the mRNA expression of PRDX4 in KYSE270 cells transfected with PRDX4 siRNA and KYSE30 cells transfected with pcDNA3.1-PRDX4. (D) CCK-8 assay of cell proliferation in KYSE270 cells transfected with PRDX4 siRNA. (E) Colony formation assay of the colony-forming ability of KYSE270 cells transfected with PRDX4 siRNA. (F) Statistical analysis of the number of colonies formed in KYSE270 cells transfected with PRDX4 siRNA. (G) CCK-8 assay of cell proliferation in KYSE30 cells transfected with pcDNA3.1-PRDX4. (H) Colony formation assay of the colony-forming ability of KYSE30 cells transfected with pcDNA3.1-PRDX4. (I) Statistical analysis of the number of colonies formed in KYSE30 cells transfected with pcDNA3.1-PRDX4. (J) EdU staining assay of EdU-positive cells in KYSE270 cells transfected with PRDX4 siRNA. Scale bar, 100 µm. (K) EdU staining assay of EdU-positive cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. Scale bar, 100 µm. (L) Statistical analysis of the number of EdU-positive cells in KYSE270 cells transfected with PRDX4 siRNA. (M) Statistical analysis of the number of EdU-positive cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; CCK-8, Cell Counting Kit-8; EdU, 5-ethynyl-2'-deoxyuridine.

Article Snippet: In addition, a control siRNA (50 nM; cat. no. sc-37007; Santa Cruz Biotechnology, Inc.) and PRDX4 siRNA (50 nM; cat. no. sc-40835; Santa Cruz Biotechnology, Inc.) were transfected into KYSE270 cells using Lipo8000 TM Transfection Reagent according to the manufacturer's instructions for 48 h at 37 ̊C.

Techniques: Knockdown, Western Blot, Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Staining, Small Interfering RNA, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 downregulation suppresses cell migration and invasion in ESCC cells. (A) PRDX4 knockdown suppresses cell migration and invasion in KYSE270 cells after transfection with PRDX4 siRNA. Scale bar, 100 µm. (B) Statistical analysis of the number of migratory cells in KYSE270 cells transfected with PRDX4 siRNA. (C) Statistical analysis of the number of invasive cells in KYSE270 cells transfected with PRDX4 siRNA. (D) Western blot analysis of the expression levels of E-cadherin, N-cadherin and vimentin in KYSE270 cells transfected with PRDX4 siRNA. (E) The relative protein levels of E-cadherin, N-cadherin and vimentin in KYSE270 cells transfected with PRDX4 siRNA. (F) PRDX4 overexpression suppresses cell migration and invasion in KYSE30 cells after transfection with pcDNA3.1-PRDX4. Scale bar, 100 µm. (G) Statistical analysis of the number of migratory cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. (H) Statistical analysis of the number of invasive cells in KYSE30 cells transfected with pcDNA3.1-PRDX4. (I) Western blot analysis of the expression levels of E-cadherin, N-cadherin and vimentin in KYSE30 cells transfected with pcDNA3.1-PRDX4. (J) The relative protein levels of E-cadherin, N-cadherin and vimentin in KYSE30 cells transfected with pcDNA3.1-PRDX4. *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA.

Article Snippet: In addition, a control siRNA (50 nM; cat. no. sc-37007; Santa Cruz Biotechnology, Inc.) and PRDX4 siRNA (50 nM; cat. no. sc-40835; Santa Cruz Biotechnology, Inc.) were transfected into KYSE270 cells using Lipo8000 TM Transfection Reagent according to the manufacturer's instructions for 48 h at 37 ̊C.

Techniques: Migration, Knockdown, Transfection, Western Blot, Expressing, Over Expression, Small Interfering RNA

Journal: Biomedical Reports

Article Title: Peroxiredoxin 4 suppresses ferroptosis in esophageal squamous cell carcinoma by activating the phosphoinositide 3-kinase  signaling pathway

doi: 10.3892/br.2026.2133

Figure Lengend Snippet: PRDX4 is an important regulator of ferroptosis in ESCC cells. (A) Determination of MDA, LPO and GSH contents in KYSE270 cells after transfection with PRDX4 siRNA. (B) Western blot analysis of the protein levels of GPX4, SLC7A11 and PTGS2 in KYSE270 cells transfected with PRDX4 siRNA. (C) The relative protein levels of GPX4, SLC7A11 and PTGS2 in KYSE270 cells transfected with PRDX4 siRNA. (D) Determination of MDA, LPO and GSH contents in KYSE30 cells after transfection with pcDNA3.1-PRDX4. (E) Western blot analysis of the protein levels of GPX4, SLC7A11 and PTGS2 in KYSE30 cells transfected with pcDNA3.1-PRDX4. (F) The relative protein levels of GPX4, SLC7A11 and PTGS2 in KYSE30 cells transfected with pcDNA3.1-PRDX4. (G) Detection of the levels of MDA, LPO and GSH in the control group, PRDX4 siRNA group and PRDX4 siRNA plus Fer-1 group in KYSE270 cells. (H) Western blot analysis of the protein expression levels of GPX4, SLC7A11 and PTGS2 in the control group, PRDX4 siRNA group and PRDX4 siRNA plus Fer-1 group in KYSE270 cells. (I) The relative protein levels of GPX4, SLC7A11 and PTGS2 in the control group, PRDX4 siRNA group and PRDX4 siRNA plus Fer-1 group in KYSE270 cells. (J) Detection of the levels of MDA, LPO and GSH in the pcDNA3.1 group, pcDNA3.1-PRDX4 group and pcDNA3.1-PRDX4 plus erastin group in KYSE30 cells. (K) Western blot analysis of the protein expression levels of GPX4, SLC7A11 and PTGS2 in the pcDNA3.1 group, pcDNA3.1-PRDX4 group and pcDNA3.1-PRDX4 plus erastin group in KYSE30 cells. (L) The relative protein levels of GPX4, SLC7A11 and PTGS2 in the pcDNA3.1 group, pcDNA3.1-PRDX4 group and pcDNA3.1-PRDX4 plus erastin group in KYSE30 cells. ** P<0.01, *** P<0.001 and **** P<0.0001, indicate statistical significance. PRDX4, peroxiredoxin 4; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA; MDA, malondialdehyde; LPO, lipid peroxidation; GSH, glutathione; GPX4, glutathione peroxidase 4; SLC7A11, solute carrier family 7 member 11; PTGS2, prostaglandin-endoperoxide synthase 2; Fer-1, ferrostatin-1; ns, not significant.

Article Snippet: In addition, a control siRNA (50 nM; cat. no. sc-37007; Santa Cruz Biotechnology, Inc.) and PRDX4 siRNA (50 nM; cat. no. sc-40835; Santa Cruz Biotechnology, Inc.) were transfected into KYSE270 cells using Lipo8000 TM Transfection Reagent according to the manufacturer's instructions for 48 h at 37 ̊C.

Techniques: Transfection, Western Blot, Control, Expressing, Small Interfering RNA

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: eIF4G2 post-transcriptionally sustains γc expression via its mRNA UTRs (A–C) Analysis in primary CD4 + CD8 lo transitional thymocytes. (A) Western blot analysis of γc and IL-7Rα protein levels. (B and C) Quantitative RT-PCR analysis of Il2rg (B) and Il7r (C) mRNA levels ( n = 3, ns p > 0.05, ∗ p < 0.05). (D–H) Mechanistic dissection in 293T cells. (D) Western blot of γc protein in control and EIF4G2 knockdown 293T cells transfected with an IL2RG coding sequence construct containing its native 5′ and 3′ UTRs. (E) Corresponding IL2RG mRNA levels measured by RT-qPCR ( n = 3, ns p > 0.05) . (F and G) Assessment of γc protein stability ( n = 3, ns p > 0.05). (F) Representative western blots and (G) quantification of γc protein levels over time following cycloheximide (CHX) treatment in si-control and si- EIF4G2 293T cells ( n = 3, ns p > 0.05). (H) Western blot of γc protein in si-control and si- EIF4G2 293T cells transfected with an IL2RG CDS construct lacking UTRs. Data are representative of at least two independent experiments. Bar graphs show mean ± SEM and unpaired Students’ t test was used to perform the statistical analysis.

Article Snippet: Cells were transfected either with a non-targeting control siRNA (si-Control) or human EIF4G2 targeting siRNA (si- EIF4G2 ) (Sangon Biotech) using Lipofectamine RNAiMAX transfection reagent (Invitrogen).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Dissection, Control, Knockdown, Transfection, Sequencing, Construct