Journal: Bioactive Materials

Article Title: M2 exosomes modified by hydrogen sulfide promoted bone regeneration by moesin mediated endocytosis

doi: 10.1016/j.bioactmat.2023.08.006

Figure Lengend Snippet: H 2 S modified M2 exosomes promoted osteogenic differentiation of MSCs via moesin mediated endocytosis. (A) The top 30 significantly different proteins between M2 exosomes and H 2 S pretreated M2 exosomes. (B) The expression of moesin in M0, M2 and H 2 S pretreated-M2 macrophages, as assessed by western blot. (C) The expression of moesin in exosomes derived from M0, M2 and H 2 S pretreated-M2 macrophages, as assessed by western blot. (D) The expression of meosin in macrophages and exosomes, with or without moesin siRNA treatment. (E) Immunofluorescent staining showed the endocytosis of exosomes from macrophages treated with or without moesin siRNA. (F) The mineralized nodule formation of MSCs stimulated by control or moesin siRNA pretreated-exosomes. ( G-J ) The expression of ALP and Runx2 of MSCs stimulated by control or moesin siRNA pretreated-exosomes, as assessed by qPCR analysis and western blot. (K) The mineralized nodule formation of MSCs treated with or without moesin protein (5 μg/ml). ( L-N) The expression of ALP and Runx2 of MSCs treated with or without moesin protein, as assessed by qPCR and western blot analysis. Scale bars: 10 μm (E), 50 μm (F, K). Data are presented by mean ± SD (*P < 0.05, **P < 0.01, *** p < 0.001).

Article Snippet: For osteogenic differentiation, MSCs were seeded in 6-well dishes at a density of 5 × 10 5 cells per well, and treated with osteogenic differentiation medium (α-MEM medium containing 15% FBS, 1% penicillin/streptomycin and 50 μg/mL ascorbic acid), macrophage culture conditional medium, 50 μg/ml different exosomes, or 5 μg/ml moesin protein (aa 1–346, His Tag, Sino Biological, China).

Techniques: Modification, Expressing, Western Blot, Derivative Assay, Staining

Journal: Bioactive Materials

Article Title: M2 exosomes modified by hydrogen sulfide promoted bone regeneration by moesin mediated endocytosis

doi: 10.1016/j.bioactmat.2023.08.006

Figure Lengend Snippet: H 2 S modified M2 exosomes promoted osteogenic differentiation of MSCs through β-catenin signaling pathway. (A) The expression of β-catenin and active β-catenin in control MSCs, MSCs treated by exosomes derived from M0, M2 and H 2 S pretreated-M2 macrophages, as assessed by western blot. (B) The expression of β-catenin and active β-catenin in control MSCs or moesin siRNA pretreated-exosomes treated MSCs. (C) The protein levels of p-β-catenin, active-β-catenin, and β-catenin in control and moesin groups treatment with CHX (20 μg/mL) for 10h, then with MG132 (10 μmol/L) for 2h. ( D )The expression of active-β-catenin in the cell cytoplasm and nucleus fraction with or without meosin treatment. (E) The mineralized nodule formation in control, stimulated by moesin with or without XAV-939 (3 μM) treatment groups. ( F–H ) The expression of ALP and Runx2 of MSCs in control, stimulated by moesin with or without XAV-939 treatment groups, as assessed by qPCR analysis and western blot. (I) The schema showed that H 2 S promoted M2 macrophages polarization with enhancing the level of exosomal protein moesin, which facilitated the uptake of exosomes by MSCs. Thus, the exosomes, from H 2 S pretreated M2 macrophages, promoted bone regeneration of MSCs via activating β-catenin signaling pathway. Scale bars: 50 μm Data are presented by mean ± SD (*P < 0.05, **P < 0.01, *** p < 0.001).

Article Snippet: For osteogenic differentiation, MSCs were seeded in 6-well dishes at a density of 5 × 10 5 cells per well, and treated with osteogenic differentiation medium (α-MEM medium containing 15% FBS, 1% penicillin/streptomycin and 50 μg/mL ascorbic acid), macrophage culture conditional medium, 50 μg/ml different exosomes, or 5 μg/ml moesin protein (aa 1–346, His Tag, Sino Biological, China).

Techniques: Modification, Expressing, Derivative Assay, Western Blot

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Quantification of IST amplitude in bRG cells following treatment with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M) in cortical organoids (N=3 organoid batches, 329 bRG cells, weeks 9-12). (B) Live imaging of mitotic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (week 8-11). shRNA plasmids co-express GFP. (C) Live imaging of mitotic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (pcw 16-20). (D) Live imaging of in vitro interphasic human bRG cells expressing control or LIS1 shRNA constructs. (E) Quantification of IST amplitude in in vitro interphasic human bRG cells expressing control or LIS1 shRNA constructs (N=3 experiments, 520 bRG cells). (F) Quantification of IST amplitude in in vitro interphasic human bRG cells expressing control or KASH constructs, in the presence of DMSO or blebbistatin (10 μ M) (N=3 experiments, 1198 bRG cells). Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. **p<0,01; ***p<0,001, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Imaging, Expressing, Control, shRNA, Construct, In Vitro, Two Tailed Test

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Live imaging of interphasic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (week 8-11). shRNA plasmids co-express GFP. (B) Live imaging of interphasic human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (pcw 16-18). (C) Quantification of IST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (N=3 organoid batches, 899 bRG cells, week 8-11). Two independent shRNA plasmids were used for each knockdown. (D) Quantification of MST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human cortical organoids (N=3 organoid batches, 899 bRG cells, week 8-11). (E) Quantification of IST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (N=3 fetal samples, 385 bRG cells, pcw 16-20). (F) Quantification of MST amplitude in human bRG cells expressing control, DYNC1H1 or LIS1 shRNA constructs in human fetal tissue (N=3 fetal samples, 385 bRG cells, pcw 16-20). (G) Live imaging of interphasic human bRG cells expressing GFP in control cortical organoids and two different patient-derived LIS1-mutated organoids (week 8-11). (H) Quantification of IST amplitude in control cortical organoids and two different patient-derived LIS1-mutated organoids (N=3 organoid batches, 397 bRG cells, week 8-11). (I) Quantification of MST amplitude in control cortical organoids and two different patient-derived LIS1-mutated organoids (N=3 organoid batches, 397 bRG cells, week 8-11). (J) Immunostaining for SOX2 and Nesprin-2 in cortical organoids expressing GFP or the KASH dominant negative together with GFP (week 9). Red arrows indicate nuclear envelope of construct-expressing cells. (K) Live imaging of interphasic human bRG cells expressing control or KASH constructs in human cortical organoids (week 8). KASH plasmid co-expresses GFP. (L) Live imaging of interphasic human bRG cells expressing control or KASH constructs in human fetal tissue (pcw 16). KASH plasmid co-expresses GFP. (M) Quantification of IST amplitude in human bRG cells expressing control or KASH constructs in human cortical organoids (N=3 organoid batches, weeks 8-11, 201 bRG cells). (N) Quantification of MST amplitude in human bRG cells expressing control or KASH constructs in human cortical organoids (N=3 organoid batches, weeks 8-11, 201 bRG cells). (O) Quantification of IST amplitude in human bRG cells expressing control or KASH constructs in human fetal tissue (N=2 fetal samples, pcw 16-18, 40 bRG cells). (P) Quantification of MST amplitude in human bRG cells expressing control or KASH constructs in human fetal tissue (N=2 fetal samples, pcw 16-18, 40 bRG cells). Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. **p<0,01; ****p<0,0001, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Imaging, Expressing, Control, shRNA, Construct, Knockdown, Derivative Assay, Immunostaining, Dominant Negative Mutation, Plasmid Preparation, Two Tailed Test

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Immunostaining for SOX2 and p-ERM in mitotic bRG cell from week 8 cortical organoid. (B) Live imaging of in vitro mitotic human bRG cells expressing control, Moesin, Radixin, Ezrin or Vimentin shRNA constructs. (C) Quantification of IST amplitude in in vitro interphasic human bRG cells expressing control, Moesin, Radixin, Ezrin or Vimentin shRNA constructs. Two independent shRNA plasmids were used for each knockdown (N=7 control samples and N=3 sample per shRNA condition, 522 bRG cells). (D) Live imaging of mitotic human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human cortical organoids (week 9). (E) Live imaging of mitotic human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human fetal tissue (pcw 20). (F) Quantification of MST amplitude in human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human cortical organoids (N=3 organoid batches, 453 bRG cells, weeks 8-12). (G) Quantification of IST amplitude in human bRG cells expressing control, Moesin or Vimentin shRNA constructs in human cortical organoids (N=3 organoid batches, 453 bRG cells, weeks 8-12). (H) Quantification of MST amplitude in human bRG cells expressing expressing control, Moesin or Vimentin shRNA constructs in human fetal tissue (N=3 fetal samples, 529 bRG cells, pcw 14-20). (I) Quantification of IST amplitude in human bRG cells expressing expressing control, Moesin or Vimentin shRNA constructs in human fetal tissue N=3 fetal samples, 529 bRG cells, pcw 14-20). (J) Schematic representation of the molecular mechanisms driving IST and MST. Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. *p<0,05; **p<0,01, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Immunostaining, Imaging, In Vitro, Expressing, Control, shRNA, Construct, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: Two independent translocation modes drive neural stem cell dissemination into the human fetal cortex

doi: 10.1101/2025.01.08.631865

Figure Lengend Snippet: (A) Live imaging of an in vitro GBM cell (line U3123) performing IST. (B) Live imaging of an in vitro GBM cell (line U3123) performing MST. (C) Quantification of the fraction of cells performing IST, in 9 GBM lines and compared to in vitro bRG cells (N=3 replicates per line, 1130 cells). (D) Quantification of the fraction of cells performing MST, in 9 GBM lines and compared to in vitro bRG cells (N=3 replicates per line, 1130 cells). (E) Immunostaining for bRG markers SOX2 and HOPX in U3123 GBM line. (F) Live imaging of U3123 GBM line during interphase, treated with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M). (G) Quantification of IST amplitude following treatment with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M) in U3123 GBM line (N=3 experiments, 274 GBM cells). (H) Live imaging of U3123 GBM line during mitosis, treated with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M). (I) Quantification of MST amplitude following treatment with DMSO, nocodazole (1 μ M) or blebbistatin (10 μ M) in U3123 GBM line (N=3 experiments, 274 GBM cells). (J) Live imaging of interphasic U3123 GBM cells expressing control, LIS1 shRNA or KASH dominant negative constructs. (K) Quantification of IST amplitude in U3123 GBM cells expressing control or LIS1 shRNA constructs. (N=3 experiments, 359 GBM cells). (L) Quantification of IST amplitude in U3123 GBM cells expressing control or KASH dominant negative constructs (N=3 experiments, 300 GBM cells). (M) Live imaging of mitotic U3123 GBM cells expressing control, Moesin or Vimentin shRNA constructs. (N) Quantification of MST amplitude U3123 GBM cells expressing control, Moesin or Vimentin shRNA (N=3 experiments, 149 GBM cells). Yellow arrowheads indicate bRG cell soma, and green and red arrowheads indicate daughter cells. Data are presented as mean values +/− SD. Scale bar = 20 µm. All live imaging montages are in hours:minutes. **p<0,01; ***p<0,001; ****p<0,0001, ns: non-significant by two-tailed unpaired t-tests.

Article Snippet: The following plasmids were used in this study: MSCV-IRES-GFP (Tannishtha Reya, Addgene 20672); VSVG (a gift from P. Benaroch), Human EZR shRNA (TF308420, Origene), Human STK10 shRNA (TF320540, Origene), Human SLK shRNA (TG320620, Origene), Human DYNC1H1 shRNA (TL313335, Origene), Human RDX shRNA (TL309884, Origene), Human MSN shRNA (TL311375, Origene), Human ECT2 shRNA (TL304854, Origene), Human VIM shRNA (TL308419, Origene), Human PAFAH1B1 (LIS1) shRNA (TL310628, Origene), Dominant Negative KASH.

Techniques: Imaging, In Vitro, Immunostaining, Expressing, Control, shRNA, Dominant Negative Mutation, Construct, Two Tailed Test

Journal: Journal of Clinical Pathology

Article Title: Microbial infections in eight genomic subtypes of chronic fatigue syndrome/myalgic encephalomyelitis

doi: 10.1136/jcp.2009.072561

Figure Lengend Snippet: CFS/ME-associated genes and transcription factors in patients with CFS/ME, Q-fever-associated CFS/ME and endogenous depression

Article Snippet: MSN , NM_002444 , Hs00792607_mH , 4.85 , 0.0016 , 7.49 , 0.002 , 1.35 , 0.962.

Techniques:

Journal: Journal of Clinical Pathology

Article Title: Microbial infections in eight genomic subtypes of chronic fatigue syndrome/myalgic encephalomyelitis

doi: 10.1136/jcp.2009.072561

Figure Lengend Snippet: Fold-difference values for 88 genes in each of eight subtypes (A–H) in 114 subtyped patients with chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME). Genes without values for the subtypes are those for which there was missing data for one or more subtypes. Bold type indicates genes targeted by existing drugs and those CFS/ME subtypes in which fold-difference values of 1.5 were found

Article Snippet: MSN , NM_002444 , Hs00792607_mH , 4.85 , 0.0016 , 7.49 , 0.002 , 1.35 , 0.962.

Techniques: