Journal: Cell Death & Disease

Article Title: Cancer-associated SF3B1 mutation suppresses DNA repair by disrupting the organization of nuclear actin network

doi: 10.1038/s41419-026-08569-5

Figure Lengend Snippet: A A Venn diagram showing the numbers of circATP9B-binding proteins in Hela and HEK293T cells that were pulled down by circATP9B probe and identified by LC-MS/MS. B Peptide counts from LC-MS/MS analysis of some circATP9B-binding proteins that were identified in both Hela and HEK293T cells. C RNA pulldown using circATP9B and control probes (Ctrl probe) and western blot analysis to confirm the interaction between circATP9B and MYH9 in Hela cells. D RNA immunoprecipitation analysis using MYH9 antibody and IgG as control to confirm the interaction between circATP9B and MYH9 in Hela cells. GAPDH was used as control. E Representative immunofluorescence images of the localization of circATP9B (red) and MYH9 (green) in Hela cells. Nuclei were stained with DAPI (blue), scale bar = 10 μm. F Schematic diagram of full-length and truncated MYH9 protein. G RIP assays with anti-Flag antibody in HEK293T cells transfected with MYH9 constructs. Co-precipitated RNAs were analyzed by qRT-PCR. H RNA pulldown assays using biotin-labeled circATP9B probe and Ctrl probe in HEK293T cells expressing full-length or truncated MYH9, co-precipitated proteins were detected by immunoblots using anti-Flag antibodies. I Schematic illustration of the potential binding sequences of MYH9 in circATP9B and circATP9B expression constructs with indicated deletion were shown. J RNA pulldown assays using biotin-labeled Ctrl probe and circATP9B probe in HEK293T cells transfected with indicated circATP9B expression constructs. Co-precipitated proteins were detected by immunoblots using anti-MYH9 antibodies. K RIP assays with anti-MYH9 antibody in HEK293T cells transfected with indicated circATP9B expression constructs. Co-precipitated RNAs were analyzed by qRT-PCR. L Western blots to show the expression of MYH9 in Hela cells with or without circATP9B overexpression and treated with 100 μM cycloheximide at the indicated times. GAPDH was used as control. M Quantification of the relative expression of MYH9 protein through calculating the intensities of western blot bands shown in ( L ). N Western blot to show ubiquitination level of MYH9 in circATP9B overexpression or control Hela cells treated with 20 μM MG132 for 12 h. Data are expressed as the mean ± SD ( D , G , K , M ), one-way ANOVA ( D , G , K ) and two-way ANOVA with multiple comparisons ( M ). ns, non-significant, *** p < 0.001.

Article Snippet: For γH2AX (Millipore, cat#05-636), Flag (Proteintech, cat#20543-1-AP), MYH9 (Proteintech, cat#60233-1-Ig), and HP1α (CST, cat#2616) antibody, cells were fixed in 4% paraformaldehyde for 20 min and permeabilized with 0.1% Triton X-100 at room temperature for 20 min. For the RAD51 (GeneTex, cat#100469) antibody, immunofluorescence staining was performed as previously described [ ].

Techniques: Binding Assay, Liquid Chromatography with Mass Spectroscopy, Control, Western Blot, RNA Immunoprecipitation, Immunofluorescence, Staining, Transfection, Construct, Quantitative RT-PCR, Labeling, Expressing, Over Expression, Ubiquitin Proteomics

Journal: Cell Death & Disease

Article Title: Cancer-associated SF3B1 mutation suppresses DNA repair by disrupting the organization of nuclear actin network

doi: 10.1038/s41419-026-08569-5

Figure Lengend Snippet: A Representative immunofluorescence images of γH2AX foci in Hela cells transfected with control siRNA (siNC) or MYH9 siRNA (siMYH9) at indicated times after irradiation were shown. γH2AX was stained with red. Scale bar = 10 μm. B Quantification of γH2AX foci at different times after irradiation (50 cells per group). C Representative images of nuclear actin filaments in siNC and siMYH9 Hela cells and treated with irradiation. Scale bar = 10 μm. D Quantification of the intensities of nuclear actin filaments in siNC and siMYH9 Hela cells images as shown in ( C ) ( n ≥ 15). E Representative images of immunofluorescence to show the localization of γH2AX (red) and Flag-NLS-MYH9 (green) after irradiation. Nuclei were stained with DAPI (blue), scale bar = 10 μm. F Western blot analysis of proteins immunoprecipitated with anti-Flag antibody in Hela cells transfected with Flag-MYH9 or control vectors and treated or non-treated with irradiation. Data are expressed as the mean ± SEM ( B ) or SD ( D ), unpaired two-tailed Student’s t -test ( B ) and two-way ANOVA with multiple comparisons ( D ). ns non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For γH2AX (Millipore, cat#05-636), Flag (Proteintech, cat#20543-1-AP), MYH9 (Proteintech, cat#60233-1-Ig), and HP1α (CST, cat#2616) antibody, cells were fixed in 4% paraformaldehyde for 20 min and permeabilized with 0.1% Triton X-100 at room temperature for 20 min. For the RAD51 (GeneTex, cat#100469) antibody, immunofluorescence staining was performed as previously described [ ].

Techniques: Immunofluorescence, Transfection, Control, Irradiation, Staining, Western Blot, Immunoprecipitation, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Cancer-associated SF3B1 mutation suppresses DNA repair by disrupting the organization of nuclear actin network

doi: 10.1038/s41419-026-08569-5

Figure Lengend Snippet: A Representative immunofluorescence images of γH2AX in Hela cells with control, circATP9B overexpression (circATP9B OE), circATP9B and MYH9 overexpression (circATP9B OE + MYH9) at indicated times after 4 Gy irradiation. γH2AX was stained with red, scale bar = 10 μm. B Quantification of γH2AX foci as shown in ( A ) after irradiation (50 cells per group). C Representative images of nuclear actin filaments in control, circATP9B OE, circATP9B OE + MYH9 Hela cells at indicated times after irradiation. D Quantification of fluorescence intensities from nuclear actin filaments ( n ≥ 15). Data are expressed as the mean ± SEM ( B ) or SD ( D ), unpaired two-tailed Student’s t -test ( B ) and two-way ANOVA with multiple comparisons ( D ). ns, non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For γH2AX (Millipore, cat#05-636), Flag (Proteintech, cat#20543-1-AP), MYH9 (Proteintech, cat#60233-1-Ig), and HP1α (CST, cat#2616) antibody, cells were fixed in 4% paraformaldehyde for 20 min and permeabilized with 0.1% Triton X-100 at room temperature for 20 min. For the RAD51 (GeneTex, cat#100469) antibody, immunofluorescence staining was performed as previously described [ ].

Techniques: Immunofluorescence, Control, Over Expression, Irradiation, Staining, Fluorescence, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Cancer-associated SF3B1 mutation suppresses DNA repair by disrupting the organization of nuclear actin network

doi: 10.1038/s41419-026-08569-5

Figure Lengend Snippet: A Representative images of mCherry-RAD52 foci traces in control, circATP9B OE, circATP9B OE + MYH9 cells over 100 min after 4 Gy irradiation. Scale bar = 2 μm. B–F MSD ( B ), distance ( C ), velocity ( D ), sizes ( E ), and clustering events ( F ) of mCherry-RAD52 foci in control, circATP9B OE, circATP9B OE + MYH9 cells. 2207 foci from 12 nuclei of control,1621 foci from 14 nuclei of circATP9B OE, 1747 foci from 12 nuclei of circATP9B OE + MYH9. Δt , time intervals. G , H Numbers of γH2AX foci ( G ) and micronuclei associated with HP1α ( H ) in control, circATP9B OE, circATP9B OE + MYH9 cells at 48 h after 4 Gy irradiation (50 cells per group for γH2AX foci, n > 110 cells per group for micronuclei). I Images of tumors obtained from BALB/c nude mice implanted with control or circATP9B OE cells and treated or non-treated with olaparib. J , K Quantification of weight ( J ) and volume ( K ) of tumors as shown in ( I ) ( n = 6). Data are expressed as the mean ± SEM ( B–H ) or SD ( J , K ), one-way ANOVA ( C–H , J ) and two-way ANOVA with multiple comparisons (B,K). ns, non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For γH2AX (Millipore, cat#05-636), Flag (Proteintech, cat#20543-1-AP), MYH9 (Proteintech, cat#60233-1-Ig), and HP1α (CST, cat#2616) antibody, cells were fixed in 4% paraformaldehyde for 20 min and permeabilized with 0.1% Triton X-100 at room temperature for 20 min. For the RAD51 (GeneTex, cat#100469) antibody, immunofluorescence staining was performed as previously described [ ].

Techniques: Control, Irradiation

Journal: bioRxiv

Article Title: MYH9, a cytosolic myosin protein, binds to dengue virus 3’UTR and facilitates replication and cellular entry

doi: 10.64898/2026.03.02.708980

Figure Lengend Snippet: (A) Co-localization assay of MYH9 with Calnexin in A549 cells after dengue virus infection. Images (i-v) uninfected cells, (vi-ix) dengue infected cells (Arrows represent the MYH9 and calnexin, (x) Enlarged (B) (i) Represents fluorescence intensity plot of uninfected cells showing unmerged green (MYH9) and red (calnexin) colours, (ii) Merged green (MYH9) and red (calnexin) colours in DENV infected cells. (C) Co-localization assay of MYH9 and dsRNA in A549 cells after dengue virus infection. Images (i-v) Represent uninfected cells, (vi-x) Dengue infected cells (Arrows represent the MYH9 and dsRNA, (x) Enlarged. (D) (i) Represents fluorescence intensity plot of uninfected cells showing unmerged green (MYH9) and red (dsRNA) colours, (ii) Merged green (MYH9) and red (dsRNA) colours in DENV infected cells. (E) Representative western blot analysis of dengue virus protein NS2BNS3 upon silencing of Myh9 mRNA. (F) The bar graph representation of the above experiment. (G) Representative gel picture for RT-PCR analysis of dengue genome (5’end) using the cell culture supernatants under the anti-MYH9 siRNA expression. (H) Bar graph representation of RT-PCR of dengue virus gene segment in siRNA transfected cell supernatant.

Article Snippet: The sources and used dilutions of the antibodies: Polyclonal anti-MYH9 (Proteintech, 1:10000), anti-actin (Santa Cruz, 1:1000), anti-GAPDH (Proteintech, 1:10000), anti-NS2BNS3 (in-house raised, 1:500), anti-calnexin (Merck, 1:200), anti-dsRNA (Merck, 1:200).

Techniques: Virus, Infection, Fluorescence, Western Blot, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Expressing, Transfection

Journal: bioRxiv

Article Title: MYH9, a cytosolic myosin protein, binds to dengue virus 3’UTR and facilitates replication and cellular entry

doi: 10.64898/2026.03.02.708980

Figure Lengend Snippet: (A) Optimization of siRNA concentration for efficient knockdown of the gene at 50 nM, 100 nM, and 200 nM. Anti-MYH9 antibodies were used to detect the MYH9 levels. (B) Graphical representation of MYH9 levels under different concentrations of siRNA. (C) Graphical representation of Myh9 at 200nM concentration relative to scRNA and NC.

Article Snippet: The sources and used dilutions of the antibodies: Polyclonal anti-MYH9 (Proteintech, 1:10000), anti-actin (Santa Cruz, 1:1000), anti-GAPDH (Proteintech, 1:10000), anti-NS2BNS3 (in-house raised, 1:500), anti-calnexin (Merck, 1:200), anti-dsRNA (Merck, 1:200).

Techniques: Concentration Assay, Knockdown