Journal: EBioMedicine

Article Title: Circulating exosomes and gut microbiome induced insulin resistance in mice exposed to intermittent hypoxia: Effects of physical activity

doi: 10.1016/j.ebiom.2021.103208

Figure Lengend Snippet: Effects on insulin sensitivity on naïve adipocytes by plasma exosomes derived from mice exposed to IH or RA for 6 weeks with and without PA. (a) Representative Western blots of pAKT and tAKT after treatment with plasma exosomes followed by 5 nM of exogenous insulin for 30 min. (b) Mean pAKT/tAKT ratios, n = 8 per experimental conditions. (c) Effects of plasma exosomes on CD11c expression by naïve macrophage (RAW 264.7) cells using flow cytometry. An equal number of exosomes (20 × 10 6) ) was used from each condition. * Indicates statistical significance of IH vs. IH+PA, p < 0.05, n = 8/condition. Two-way of variance (ANOVA) for non-repeated measures was used. NS indicates not significant.

Article Snippet: The mouse macrophage cell line RAW cells, RAW 264.7, (ATCC® TIB-7) (40 × 10 4 ) was seeded in 24-well plates in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum, 1% 100 mM sodium pyruvate, 1% of 10,000 U/mL penicillin and 10,000 μg/mL streptomycin and maintained in 37 °C humidified incubator containing 5% CO 2 .

Techniques: Clinical Proteomics, Derivative Assay, Western Blot, Expressing, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: An apoA-I mimetic peptide increases LCAT activity in mice through increasing HDL concentration

doi:

Figure Lengend Snippet: ABCA1 dependent cholesterol efflux assay. The assay details are described in Materials and Methods. A, ABCA1 expression levels in RAW cells were determined by western blot with anti-mouse ABCA1 antibody. Very low basal expression levels of ABCA1 are dramatically increased by the addition of cAMP. B, apoA-I-stimulated cholesterol efflux from macrophage is ABCA1 level dependent. C, apoA-I and its formulated particles show activity in stimulating cholesterol efflux through the ABCA1 transporter, providing EC 50 values of 101 nM and 103 nM, respectively. D, D-4F and its formulated particles show activity in stimulating cholesterol efflux through the ABCA1 transporter, providing EC 50 values of 2.2 μM and 2.8 μM, respectively. F, both size and discoidal shape of D-4F particles made with Dipalmitoylphosphatidylcholine ( DPPC ) (D4F:DPPC = 1:10) were confirmed by electron microscope (Magnification 45000, disk size; 6 x 12 nm).

Article Snippet: Mouse macrophage cells (RAW 264.7, ATCC) were cultured in Dulbecco's Modified Eagle Medium (Invitrogen 12491) with 10% FCS and 1% Pen Strep Glutamine (GIBCO 10378) in T225 flasks to a confluency of ~80%.

Techniques: Expressing, Western Blot, Activity Assay, Microscopy

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) with anti-VEGFR2 antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Raw images were converted into Imaris-compatible files using the Imaris File Converter software.

Techniques: Mutagenesis, Generated, Software, Injection, Isolation, Immunoprecipitation, Control, Western Blot, Marker, Staining, Fluorescence

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), Brain tissue extracts were prepared as in Methods from twelve PS1 FAD patients each carrying a different PS1 mutation, and twelve non-demented controls described in Methods. Left: VEGFR2 dimers and monomers were detected in brain extracts on WBs using anti-VEGFR2 antibody D5B1. Representative gels with control (C1-4) or FAD samples (FAD1-4) expressing mutants P264L, A260V, N135S and P242H respectively are shown. Vinculin: loading control. Right: Graph shows the fold change in VEGFR2 dimer to monomer ratio of FAD and control samples. (B), Brain tissue extract from control and PS1 FAD patient brains described in 7A were prepared and IPed with anti-endoglin antibody (ab252345) or IgG as in Methods. Upper panel: VEGFR2 co-IPed with endoglin was detected on WBs using an anti-VEGFR2 antibody as in 7A. Lower panel: Input samples are shown. Representative gel with control samples (C1, C2) and FAD samples (FAD1, FAD2) expressing mutants A260V and P264L respectively is shown. β-actin: loading control. Right: Graph shows relative levels of VEGFR2 co-precipitated with endoglin. (C), Brain sections from control and PS1 FAD patients were prepared as in Methods and stained for Col IV as in 1A. Upper: Representative images show brain vessels in either PS1 FAD or control (CT) brain sections. Scale bar: 80μm. Lower: Graph shows total vessel length density in PS1 FAD and CT brains quantified with Imaris software as in 1A. A-C , bars represent Mean ± S.E. For statistical analysis, unpaired t-test was performed. *p < 0.05, **p<0.01 and ***p<0.001.

Article Snippet: Raw images were converted into Imaris-compatible files using the Imaris File Converter software.

Techniques: Mutagenesis, Control, Expressing, Staining, Software

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: LRRK2 colocalized with LC3 on lysosomal single membranes. (A) Fluorescence images of endogenous LRRK2 and LC3 in cells in the absence of any stresses (upper panels), treated with CQ (middle panels) or with zymozan (lower panels). Scale bars, 10 and 1 μm (insets). (B) Percentages of LRRK2-positive phagosomes in LC3-positive and -negative phagosomes. Data represent mean ± SEM ( N = 3 independent experiments). The difference was analyzed using an unpaired two-tailed t test. (C) Fluorescence images of RAW264.7 cells cultured in amino acid–depleted medium for 90 min. (D) Fluorescence images of RAW264.7 cells treated with LLOMe or CQ. Arrowheads indicate Gal3-positive and LRRK2-negative lysosomes and arrows indicate LRRK2-positive and Gal3-negative lysosomes. Scale bar, 10 μm. (E) Percentage of cells harboring LRRK2-positive or Gal3-positive vacuoles over time under CQ or LLOMe treatment, as shown in D. Data represent mean ± SEM ( N = 4 independent experiments, 30–74 cells were analyzed in each experiment). (F) CLEM analysis of LRRK2-LC3 double-positive structures. Scale bars, 10 μm, 1 μm (inset-1, inset-2). Arrows: LRRK2-LC3 double-positive membrane, black arrowhead: autophagosome, white arrowhead: nuclear envelope.

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Fluorescence, Two Tailed Test, Cell Culture, Membrane

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Detailed analyses on LRRK2 localization. (A) Confirmation of the specificity of LRRK2 immunostaining. Immunofluorescence signal of LRRK2 stained with MJFF2 antibody was not detected in Lrrk2 KO RAW264.7 cells in the presence or absence of CQ treatment. Scale bars, 10 μm. (B and C) Two representative CLEM images, in addition to , showing colocalization of GFP-LC3 and mCherry-LRRK2 on lysosomal single membranes under CQ treatment. Scale bars, 5 μm (B), 10 μm (C), 1 μm (inset-1 in B and inset in C), 500 nm (inset-2 in B). Arrows: LRRK2-LC3 double-positive membranes, black arrowhead in B: autolysosome, white arrowheads: nuclear envelope.

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Immunostaining, Immunofluorescence, Staining

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: The ATG8 conjugation system regulates lysosomal and phagosomal targeting of LRRK2 independently of the autophagy initiation complex. (A) Levels of lipidated LC3 (LC3-II) in RAW264.7 cells transfected with nontarget or LRRK2 siRNA and treated with CQ. (B) Quantification of LC3-II in cells, as shown in A. Data represent mean ± SEM ( N = 4 independent experiments). The difference was analyzed using one-way ANOVA with Tukey’s test. ns, not significant. (C and D) Fluorescence images of endogenous LRRK2 in RAW264.7 cells transfected with the indicated siRNAs and treated with CQ (C) or zymosan (D). Arrows in C indicate LRRK2-positive lysosomes. Scale bars, 10 and 1 μm (inset). (E and F) Percentage of cells harboring LRRK2-positive lysosomes (E), as shown in C, or LRRK2-positive phagosomes (F), as shown in D. Data represent mean ± SEM ( N = 3 [E] and N = 4 [F] independent experiments). The difference was analyzed using one-way ANOVA with Dunnet’s test. (G and H) Fluorescence images of endogenous LRRK2 in RAW264.7 cells transfected with nontarget or FIP200 siRNA and treated with CQ (G) or zymosan (H). Scale bars, 10 μm. (I and J) Percentages of cells harboring LRRK2-positive lysosomes (I) or phagosomes (J) as shown in G and H, respectively. Data represent mean ± SEM ( N = 10 independent experiments). The difference was analyzed using unpaired two-tailed t test. ns, not significant. Source data are available for this figure: .

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Conjugation Assay, Transfection, Fluorescence, Two Tailed Test

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Confirmation of ATG5-dependent recruitment of LRRK2 to lysosomes upon CQ treatment. (A) MCC analysis showing the ratio of LRRK2 on LAMP1-positive area in CQ-treated and untreated RAW264.7 cells. The difference was analyzed using one-way ANOVA with Tukey’s test. (B) Biochemical isolation of lysosomes from RAW264.7 cells showing the enrichment of LRRK2 in lysosomal fraction upon treatment with CQ but not with siATG5. Source data are available for this figure: .

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Isolation

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Analyses of knockdown effects of ATG13 and FIP200. (A) Fluorescence images of endogenous LRRK2 in RAW264.7 cells transfected with nontarget or ATG13 siRNA and treated with CQ. Arrows indicate LRRK2-positive lysosomes. Scale bar, 10 μm. (B) Percentages of cells harboring LRRK2-positive lysosomes as shown in A. Data represent mean ± SEM ( N = 3 independent experiments). The difference was analyzed using one-way ANOVA with Dunnet’s test. ns, not significant. (C and D) Immunoblot pictures showing efficient knockdown of FIP200 (C) or ATG13 (D) as well as resultant decrease of starvation-induced autophagy as assessed by LC3 lipidation in RAW264.7 cells treated with siFIP200 or siATG13. Source data are available for this figure: .

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Knockdown, Fluorescence, Transfection, Western Blot

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Analyses on the possibility of LRRK2 recruitment via ATG8 or Rab29. (A and B) Lack of binding between LRRK2 and ATG8 family proteins. Immunoprecipitation (IP) analysis of HEK293 cells overexpressing 3×FLAG-taggted LC3A/B/C (A) or GABARAP/GABARAPL1/GABARAPL2 (B) together with GFP-tagged LRRK2 or p62. Pulldown using GFP-trap resulted in the precipitation of LC3 with p62 but not with LRRK2. (C) Fluorescence images of endogenous Rab29 in RAW264.7 cells transfected with siATG16L1 or siRab29 (positive control) and treated with (right) or without (left) CQ. Scale bar, 10 μm. (D) Percentage of cells harboring Rab29-positive lysosomes, as shown in C. Data represent mean ± SEM ( N = 3 independent experiments). The difference was analyzed using one-way ANOVA with Dunnet’s test. Source data are available for this figure: .

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Binding Assay, Immunoprecipitation, Fluorescence, Transfection, Positive Control

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Factors involved in LC3-assosiated phagocytosis and the V-ATPase–ATG16L1 axis are required for the targeting of LRRK2. (A) Fluorescence images of LRRK2 in zymosan-treated RAW264.7 cells transfected with the indicated siRNAs. Scale bars, 10 and 1 μm (inset). (B) Percentage of LRRK2-positive phagosomes in total phagosomes, as shown in A. Data represent mean ± SEM ( N = 3 independent experiments). The differences were analyzed using one-way ANOVA with Tukey’s test. (C) Fluorescence images of LRRK2 in HEK293 cells transfected with GFP-SopF. Arrows indicate LRRK2-positive lysosomes. Scale bar, 10 μm. (D) Percentage of cells harboring LRRK2-positive lysosomes in CQ-treated HEK293 cells, as shown in C. Data represent mean ± SEM ( N = 11 independent experiments). The difference was analyzed using an unpaired two-tailed t test. (E) Fluorescence images of LRRK2 in CQ-treated BMDMs (WT or E230). Arrows indicate LRRK2-positive lysosomes. Scale bar, 10 μm. (F) Percentage of cells harboring LRRK2-positive vacuoles in CQ-treated BMDMs, as shown in E. n = 109, 146 cells, respectively. The difference was analyzed using Fisher’s exact test. (G) Fluorescence images of LRRK2 in zymosan-treated BMDMs (WT or E230). Arrows indicate LRRK2-positive phagosomes. Scale bar, 10 μm. (H) Percentage of cells harboring LRRK2-positive phagosomes in zymosan-treated BMDMs, as shown in G. n = 112, 107 cells, respectively. The difference was analyzed using Fisher’s exact test.

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Fluorescence, Transfection, Two Tailed Test

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Kinase activity of LRRK2 is regulated by the ATG8 conjugation system. (A) Levels of phosphorylated Rab10 (pThr73) and other indicated proteins in CQ-treated or untreated RAW264.7 cells transfected with the indicated siRNAs. (B) Quantification of pRab10 divided by total Rab10 in RAW264.7 cells, as shown in A. Data represent mean ± SEM ( N = 4 independent experiments). The difference was analyzed using one-way ANOVA with Tukey’s test. (C) Levels of phosphorylated Rab10 (pThr73) in CQ-treated or untreated BMDMs derived from WT and E230 mice. (D) Quantification of pRab10 divided by total Rab10 in BMDMs, as shown in C. Data represent mean ± SEM ( N = 4 independent experiments). The difference was analyzed using one-way ANOVA with Tukey’s test. (E) Levels of pRab10 in CQ-treated or untreated MEF cells. Atg16l1 KO MEF cells were infected with full-length (FL) or WD40 repeat-deficient (ΔWDR) ATG16L1. (F) Quantification of pRab10 divided by total Rab10 in MEF cells, as shown in E. Data represent mean ± SEM ( N = 3 independent experiments). The difference was analyzed using one-way ANOVA with Tukey’s test. Source data are available for this figure: .

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Activity Assay, Conjugation Assay, Transfection, Derivative Assay, Infection

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: The ATG8 conjugation system facilitates lysosome maintenance by LRRK2 independently of the autophagy initiation complex. (A) Levels of mature (mat) and intermediate (int) CatD released into culture media from RAW264.7 cells transfected with the indicated siRNAs and treated with or without CQ. α-Tubulin in cell lysate was analyzed to normalize the amount of cells in each dish. (B) Quantification of CatD released into culture media upon CQ treatment, as shown in A. Data represent mean ± SEM ( N = 4 independent experiments). The difference was statistically analyzed using one-way ANOVA with Tukey’s test. P values compared to nontarget siRNA-transfected cells are shown in the graph. (C) Levels of mature (mat) CatB released into culture media from RAW264.7 cells transfected with the indicated siRNAs and treated with or without CQ. (D) Quantification of CatB released into culture media upon CQ treatment, as shown in C. Data represent mean ± SEM ( N = 6 independent experiments). The difference was statistically analyzed using one-way ANOVA with Tukey’s test. (E) Fluorescence images of morphologies of LAMP1-positive lysosomes in RAW264.7 cells transfected with the indicated siRNAs and treated with CQ. The largest lysosome in each cell was surrounded by a broken line. Scale bar, 10 μm. (F) Quantification of the size of the most enlarged lysosomes in each CQ-treated cells, as shown in E. The average size in each experiment was calculated and statistically analyzed. Data represent mean ± SEM ( N = 4 independent experiments). The difference was analyzed using one-way ANOVA with Dunnett’s test. Source data are available for this figure: .

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Conjugation Assay, Transfection, Fluorescence

Journal: The Journal of Cell Biology

Article Title: The V-ATPase–ATG16L1 axis recruits LRRK2 to facilitate the lysosomal stress response

doi: 10.1083/jcb.202302067

Figure Lengend Snippet: Morphological analysis of lysosomes in the absence of CQ. Representative fluorescence images of LAMP1-positive lysosomes as well as LRRK2 in RAW264.7 cells transfected with the indicated siRNAs, without following CQ treatment. Scale bar, 10 μm.

Article Snippet: RAW264.7 cells and Lrrk2 KO RAW264.7 cells (American Type Culture Collection) were cultured on culture dishes for suspended cells (Sumitomo Bakelite Co.) under the same conditions as for the culture of HEK293 cells.

Techniques: Fluorescence, Transfection