negative control shrna sh nc  (Genechem)

Journal: Cell Death & Disease

Article Title: Impact of C4BPA on Muscle progenitor cell differentiation: insights for Duchenne muscular dystrophy treatment

doi: 10.1038/s41419-026-08588-2

Figure Lengend Snippet: A Diagram scheme of the indirect co-culture of C4BPA siRNA-transfected FAPs with healthy myoblasts. B Relative expression of C4BPA mRNA levels after transfection ( n = 1). C Dot plot showing the differentiation index of myoblasts after being co-cultured with transfected and non-transfected DMD FAPs and healthy FAPs. D Dot plot showing the number of nuclei per myotube of myoblasts after being co-cultured with transfected and non-transfected DMD FAPs and healthy FAPs. E % of myotubes per field measured by MHCII staining after being co-cultured with transfected and non-transfected DMD FAPs and healthy FAPs. F Representative immunostaining of the indirect co-culture system analysed at the end of the differentiation process after FAPs transfection. Data are shown as means ± SD; Statistical significance was set at P < 0.05. ** P < 0.01; *** P < 0.001.

Article Snippet: The C4BPA siRNA, 5’-GCAAGUAGAGAUUAAGACAdTdT-3’ and MISSION® siRNA Universal Negative Control were synthesized by Merck Life Science.

Techniques: Co-Culture Assay, Transfection, Expressing, Cell Culture, Staining, Immunostaining

Journal: Journal of Nanobiotechnology

Article Title: Spatial and cellular composition of lung fibrosis induced by multi-walled carbon nanotubes

doi: 10.1186/s12951-026-04135-5

Figure Lengend Snippet: In vitro validation of macrophage responses to MWCNTs. ( A ) Cell viability of MH-S cells after treatment with increasing concentrations of MWCNTs for 24 h. ( B ) Internalization of MWCNTs by MH-S cells over time observed under microscopy. ( C ) Immunofluorescence staining of γ-H2AX in MH-S cells after MWCNT exposure. ( D ) Immunofluorescence detection of macrophage polarization markers (Adgre1, iNOS, Arg1) and inflammatory-related proteins (Slamf7, GSDMD). ( E ) Immunofluorescence staining of Slamf7, TNF-α, and IL-1β in MH-S cells. ( F ) Validation of Slamf7 knockdown efficiency by siRNA in MWCNT-treated MH-S cells. ( G ) Relative mRNA level of Tnf in cells with or without Slamf7 knockdown. ELISA measurement of TNF-α ( H ) and IL-1β ( I ) levels in the supernatant of MWCNT-treated MH-S cells with or without Slamf7 knockdown. A p value of less than 0.05 is considered significant (significance is denoted as * )

Article Snippet: Cells were seeded in 6-well plates (1 × 10 6 cells/well) 24 h prior to reach 60–70% confluence. siRNAs against Slamf7 (sense: 5′-UAAGUGGAGGGCACAAGUCGT-3′; antisense: 5′-CGACUUGUGCCCUCACUUAT-3′) and scrambled negative control (NC) siRNA (Sangon Biotech, China) were complexed with RNATransMate (Sangon Biotech, China).

Techniques: In Vitro, Biomarker Discovery, Microscopy, Immunofluorescence, Staining, Knockdown, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: AUTOTAC-mediated targeted degradation of transthyretin aggregates ameliorates hereditary transthyretin amyloidosis

doi: 10.64898/2026.02.23.707350

Figure Lengend Snippet: p62-dependent autophagic delivery of TTR. ( A ) Illustration of the detailed 3D docking pose of ATC201 within the mutant TTR-p62 ZZ domain complex. The mutant TTR structure is shown in cyan, while the p62 ZZ domain is depicted in brown. ATC201 features three distinct regions: the curcumin moiety (yellow), a flexible linker (green), and the terminal portion interacting with the p62 ZZ domain (purple). ( B ) Illustration of the 2D interaction diagram of ATC201 with the mutant TTR (Chain A) and the p62 ZZ domain (Chain Z). Key interactions are highlighted in red dashed boxes. ( C-D ) Immunoblotting analysis of HeLa cells transfected with TTR V30M-MYC , subsequently treated with ATC201 (1 μM, 24 h) under siRNA-mediated knockdown of p62 and Ubb (40 nM, 48 h). ( E ) Immunostaining analysis of HeLa cells transfected with TTR V30M-MYC , subsequently treated with the indicated compounds (1 μM, 24 h). ( F ) Quantification of (E) (n = 50 cells). ( G ) Immunostaining analysis of HeLa cells transfected with TTR V30M-MYC , subsequently treated with the indicated compounds (1 μM, 24 h). ( H ) Quantification of (G) (n = 50 cells). ( I ) Immunostaining analysis of HeLa cells transfected with TTR V30M-MYC , subsequently treated with ATC201 or ATC202 (1 μM, 24 h). ( J ) Quantification of (I) (n = 50 cells). ( K ) Triton X-100-fractionation assay of HeLa cells transfected with TTR V30M-MYC , subsequently treated with ATC201 (1 μM, 24 h) in the presence or absence of HCQ (10 μM, 24 h). ( L ) CoIP assay in HEK293T cells co-transfected with p62 GFP and TTR V30M-MYC , subsequently treated with ATC201 (1 μM, 24 h) in the presence or absence of HCQ (10 μM, 24h). Scale bar represents 10μm. Data are presented as means ± SD. Each n represents an independent biological replicate. One-way ANOVA; p* < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 and ns, not significant.

Article Snippet: The following oligonucleotides and siRNAs were used: siRNA negative control (Bioneer, SN-1003), siRNA against p62 (Bioneer, 1144479), siRNA against ATE1 (Thermo Fisher Scientific, 4390824), siRNA against ATG5 (Thermo Fisher Scientific, 4392420), GAPDH primer (Cosmogenetech; Fwd: 5′-TCAACAGCGACACCCACTCC-3′, Rvs: 5′-TGAGGTCCACCACCCTGTTG-3′), CHOP primer (Cosmogenetech; Fwd: 5′-GGAGCTGGAAGCCTGGTATG-3′, Rvs: 5′-GCAGGGTCAAGAGTGGTGAA-3′), BiP primer (Cosmogenetech; Fwd: 5′-AATGACCAGAATCGCCTGAC-3′, Rvs: 5′-CGCTCCTTGAGCTTTTTGTC-3′), XBP1 primer (Cosmogenetech; Fwd: 5′-AACCAGGAGTTAAGACAGCGCTT-3′, Rvs: 5′-CTGGGTCCAAGTTGTCCAGAAT-3′), and ATF4 primer (Cosmogenetech; Fwd: 5′-ATGACCGAAATGAGCTTCCTG-3′, Rvs: 5′-GCTGGAGAACCCATGAGGT-3′).

Techniques: Mutagenesis, Western Blot, Transfection, Knockdown, Immunostaining, Fractionation, Co-Immunoprecipitation Assay