Journal: European Journal of Trauma and Emergency Surgery

Article Title: Multifaceted intestinal defense following experimental blunt abdominal trauma

doi: 10.1007/s00068-026-03145-0

Figure Lengend Snippet: Posttraumatic defensive pathway activation and cellular immune response in the murine jejunum. ( A ) Proteome profiler-based pathway analysis revealed activation of immune and defense-related mechanisms. The 15 pathways with the highest degree of regulation are shown. ( B, C ) Histological quantification in jejunal tissue demonstrated a significant increase of CD3 + T lymphocytes in the trauma group relative to controls. ( D ) Gene expression profiling of S100a8 showed a significant post-traumatic upregulation in traumatized mice compared to sham animals. ( E ) In contrast, no statistically significant alteration was observed in interleukin-6 (Il6) expression. ( F ) Plasma levels of C5a were unaltered by traumatic exposure. ( G ) C5a in bronchoalveolar lavage fluid (BALF) was significantly increased in mice 24 h after AT compared to shams. C : n = 180 per group; D-G : n = 4–5 per group. Statistical analysis was performed using the Mann-Whitney U test, with significance defined as p < 0.05. Data are presented as mean ± standard error of the mean (SEM)

Article Snippet: To measure the concentrations of C5a in plasma and BALF, a sandwich ELISA was performed using the C5a ELISA Kit Mouse Duo Set (R&D Systems, Minneapolis, USA).

Techniques: Activation Assay, Gene Expression, Expressing, Clinical Proteomics, MANN-WHITNEY

Journal: Theranostics

Article Title: C5aR1 is a master regulator in Colorectal Tumorigenesis via Immune modulation

doi: 10.7150/thno.45058

Figure Lengend Snippet: The strong correlation between complement activation and CRC. ( A-D ) The associations between the mRNA levels of C3 (A), C5 (B), C5AR1 (C), and C5AR2 (D) and the overall survival of CRC patients (n=364). ( E, F ) The effect of C3 , C5 , C5ar1 or C5ar2 deficiency on AOM/DSS-induced colorectal tumorigenesis compared to WT control. Images of the colorectum, where CRC often developed at distal (rectum) sites (E), and quantitative analysis of tumor number and mass (F). The experiment was duplicated. Scale bar in (E), 1 cm. ( G ) Pathological analysis of control or tumor tissues, including H&E, Ki-67 and C3d staining. Scale bar, 100 µm for H&E and 50 µm for IHC. ( H ) C5a concentration in colon tissue homogenates (CTHs) measured by ELISA. n≥ 7 in each group of WT, C3 -KO, C5 -KO, C5ar1 -KO, or C5ar2 -KO mice. Data are represented as mean ± SEM; ns, not significant; * P <0.05; and *** P <0.001. NC, negative control.

Article Snippet: The level of mouse C5a in colon tissue homogenates (CTHs) was measured with Mouse Complement Component C5a DuoSet ELISA kit according to the manufacturer's instructions (R&D Systems, Minneapolis, MN).

Techniques: Activation Assay, Control, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: Theranostics

Article Title: C5aR1 is a master regulator in Colorectal Tumorigenesis via Immune modulation

doi: 10.7150/thno.45058

Figure Lengend Snippet: Schematic for the mechanisms by which C5a/C5aR1 signaling initiates colorectal tumorigenesis by modulating versatile immune responses. Complement C5a/C5aR1 signaling, independent of C3 activation, recruits MDSCs into the inflamed colorectal tissues to impair CD8 + T cells, and modulates the production of a variety of cytokines/chemokines (IL-1, IL-6, IL-11, IL-17A, TNF-α, IL-9, IL-10, IL-23, IL-27, CCL2, CCL17, CXCL1/5), thus fostering AOM/DSS-induced colorectal tumorigenesis. C5aR1 inhibition by PMX205 impedes CRC tumorigenesis. ACF, aberrant crypt foci.

Article Snippet: The level of mouse C5a in colon tissue homogenates (CTHs) was measured with Mouse Complement Component C5a DuoSet ELISA kit according to the manufacturer's instructions (R&D Systems, Minneapolis, MN).

Techniques: Activation Assay, Inhibition

Journal: Cell host & microbe

Article Title: Leukotriene B4-Mediated Neutrophil Recruitment Causes Pulmonary Capillaritis during Lethal Fungal Sepsis.

doi: 10.1016/j.chom.2017.11.009

Figure Lengend Snippet: Figure 2. Rapid PMN Intravascular Chemotaxis to Sequestered C. albicans Is Complement Dependent (A) Lung C5a levels were determined using ELISA in control and C3/ animals, following i.v. C. albicans. Two-way ANOVA, Sidak’s multiple comparison. (B) Using intravital microscopy, PMN chemotactic behaviors in C3/ and anti-C5aR mAb-treated mice were observed. Images shown are 10 min after C. albicans injection. Red arrows highlight yeast not recognized by PMN. Scale bar represents 30 mm. (C) Quantification of PMN chemotaxing to pathogens in C3/ and C5a receptor-blocked mice. One-way ANOVA, Dunnett’s multiple comparison. (D) The percentage of C. albicans phagocytosed during the initial 10 min of intravenous administration in control, C3/, and anti-C5aR mAb-treated mice using pulmonary intravital microscopy. t tests were used to compare C57BL/6 and anti-C5aR mAb treated (*) or C3/ (#) mouse time points. (E and F) C. albicans CFU was quantified in organs at (E) 1 hr and (F) 24 hr after injection with 1 3 106 C. albicans in mice treated with anti-C5a receptor antibodies or isotype. (G) Clinical sepsis scores of isotype and anti-C5aR mAb antibody treated mice 24 hr after injection with 1 3 106 C. albicans. A score of 21 is the clinical endpoint for euthanization. For (A)–(F), at least three individual experiments were performed; for (G) three individual experiments were performed. Error bars represent mean ± SEM. *p < 0.05, **p < 0.01.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Ly6G (clone 1A8) BioLegend RRID: AB_2563207 (BioLegend Cat. No. 127636) Anti-CD31 (clone MEC13.3) BioLegend RRID: AB_2161030 (BioLegend Cat. No. 102515) Anti-CD45 (clone 30-F11) BioLegend RRID: AB_493532 (BioLegend Cat. No. 103121) Anti-CD49b (clone HMa2) BD Bioscience Catalog No. 558295 Anti-C5aR (CD88, clone 20/70) BioLegend RRID: AB_2067286 (BioLegend Cat. No. 135804) Purified anti-mouse Ly6G (clone 1A8) BioXCell BP0075-1 Anti-TER119 Biolegend RRID: AB_528961 (BioLegend Cat. No. 116218) Anti-human CD15 (clone W6D3) BioLegend RRID: AB_756018 (BioLegend Cat. No. 323012) Anti-human CD45 (clone 2D1) R&D Systems Catalog # FAB1430G Anti-human C5aR (CD88, clone S5/1) BioLegend RRID: AB_2259318 (BioLegend Cat. No. 344302) Bacterial and Virus Strains C. albicans (strain ATCC58716) ATCC LUMC-101 Chemicals, Peptides, and Recombinant Proteins Zymosan A (S. cerevisiae) BioParticles ThermoFisher Z23373 Escherichia coli (K-12 strain) BioParticles ThermoFisher E13231 SYTO 9 Green Fluorescent Nucleic Acid Stain ThermoFisher S34854 Compstatin Tocris Cat. No. 2585 LY293111 Cayman Chemical Company CAS N 161172-51-6 Recombinant mouse C5a R&D Systems 2150-C5-025 Critical Commercial Assays LTB4 Parameter Assay Kit R&D Systems KGE006B EasySep Mouse Biotin Positive Selection Kit STEMCELL Technologies Catalog # 18556 Mouse Complement C5a PicoKine ELISA Kit Boster Biological Technology EK0987 Polydimethylsiloxane (PDMS) microfluidic devices SynvivoBio Cat # 104001 Experimental Models: Cell Lines human lung microvascular endothelial cells HLMEC Lonza CC-2527 Experimental Models: Organisms/Strains C57BL/6J The Jackson Laboratory Stock No: 000664 LTB4R knockout (B6.129S4-Ltb4r1tm1Adl/J) The Jackson Laboratory Stock No: 008102 C3 / ; (B6;129S4-C3tm1Crr/J) Gift from Paul Kubes N/A CD11b / ; (B6.129S4-Itgamtm1Myd/J) Gift from Paul Kubes N/A CD11a / ; (B6.129S7-Itgaltm1Bll/J) Gift from Paul Kubes N/A LysM-GFP Gift from Paul Kubes N/A Ly6G-cre (C57BL/6-Ly6g(tm2621(CretdTomato)Arte)) Gift from Matthias Gunzer N/A Software and Algorithms Volocity PerkinElmer N/A Leica LAS Leica N/A Prism 7 software GraphPad V7.02 FlowJo FlowJo V10.1 e1 Cell Host & Microbe 23, 1–13.e1–e4, January 10, 2018

Techniques: Chemotaxis Assay, Enzyme-linked Immunosorbent Assay, Control, Comparison, Intravital Microscopy, Injection

Journal: Neurotoxicity Research

Article Title: The Novel C5aR Antagonist DF3016A Protects Neurons Against Ischemic Neuroinflammatory Injury

doi: 10.1007/s12640-019-00026-w

Figure Lengend Snippet: a OGD/R-UT condition dramatically reduced the C5a protein level but induced a significant increase of C5aR gene expression. b DF3016A C5aR antagonist notably reduced C5aR transcription (mean ± SEM six independent experiments). * P < 0.05, ** P < 0.01, *** P < 0.000 (Bonferroni’s post hoc test). CTR/R-UT control-untreated, OGD/R-UT OGD/R exposed neurons, CTR/R-DF control treated with 500 nM DF3016A, OGD/R-DF OGD/R exposed neurons treated with 500 nM DF3016A

Article Snippet: Quantitative measurements of cellular proteins (C5a, TNF-α, IL-1β, and IL-6) in cell culture supernatants were determined by ELISA methods using Rat IL-6, Mouse TNFα, Rat IL-1β PicoKine TM ELISA Kits (Boster Biological Technology) and Rat C5a ELISA Kit (Elabscience Biotechnology Inc.) according to the manufacturer’s instructions.

Techniques: Gene Expression, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Complement C5 activation promotes type 2 diabetic kidney disease via activating STAT3 pathway and disrupting the gut‐kidney axis

doi: 10.1111/jcmm.16157

Figure Lengend Snippet: Increased complement C5 activation in kidneys of DKD patients and db/db mice. (A) Double‐IF staining of C5a and CD31 in the kidneys of humans and mice (scale bar = 50 µm). (B) The fluorescent intensity of C5a in kidney detected by IF staining. (C) The concentration of C5a in serum and renal tissue of mice measured by ELISA. (D) Double‐IF staining of C5aR and CD31 in the kidneys of humans and mice (scale bar = 50 µm). (E) The protein level of C5a and C5aR in kidney of mice measured by Western blot. & P < .05, DKD patient vs Healthy. * P < .05, db/db or db/db + C5aRA group vs db/m; # P < .05, db/db + C5aRA group vs db/db group

Article Snippet: The C5a concentrations in mouse serum and renal tissue were assayed by using a commercial ELISA detection kit (Mouse Complement C5a ELISA Kit, ab193718, Abcam) according to the manufacturer's instructions.

Techniques: Activation Assay, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Complement C5 activation promotes type 2 diabetic kidney disease via activating STAT3 pathway and disrupting the gut‐kidney axis

doi: 10.1111/jcmm.16157

Figure Lengend Snippet: Effect of SCFAs on the C5a/C5aR‐STAT3 pathway in HRGECs. (A) Double‐IF staining of p‐STAT3 in HRGECs after different treatments as indicated (scale bar = 20 µm). (B) Quantification of the nucleus/cytoplasm ratio of p‐STAT3 in HRGECs. (C) Western blot and quantitative analysis of p‐STAT3 and STAT3 protein expression in HRGECs after different treatments as indicated (n = 3, * P < .05, control group vs other groups; # P < .05, C5a + SCFAs or C5a + C5aRA group vs C5a alone group). (D) Double‐IF staining of p‐STAT3 with CD31 or C5a in renal sections from mice (scale bar = 50 µm). (E) Western blotting analysis of p‐STAT3 and STAT3 protein in the renal tissues of mice. * P < .05, db/db or db/db + C5aRA group vs db/m group; # P < .05, db/db + C5aRA group vs db/db group

Article Snippet: The C5a concentrations in mouse serum and renal tissue were assayed by using a commercial ELISA detection kit (Mouse Complement C5a ELISA Kit, ab193718, Abcam) according to the manufacturer's instructions.

Techniques: Staining, Western Blot, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Complement C5 activation promotes type 2 diabetic kidney disease via activating STAT3 pathway and disrupting the gut‐kidney axis

doi: 10.1111/jcmm.16157

Figure Lengend Snippet: The potential mechanism of complement C5 activation‐induced renal injury in T2DM. In T2DM status, the up‐regulation of C5a activated the STAT3 pathway and thus promoted inflammatory response and further renal fibrosis in kidneys. Meanwhile, the elevated C5a caused gut microbiota dysbiosis (eg declined gut microbiota diversity) with decreased SCFAs production, which impaired the anti‐inflammatory roles of endogenous SCFAs. Conversely, C5aR blockade could attenuate the renal dysfunction through suppressing the STAT3 pathway and restoring the gut‐kidney axis

Article Snippet: The C5a concentrations in mouse serum and renal tissue were assayed by using a commercial ELISA detection kit (Mouse Complement C5a ELISA Kit, ab193718, Abcam) according to the manufacturer's instructions.

Techniques: Activation Assay

Journal: Inflammation

Article Title: Red Blood Cell-Derived Exosomes Deliver Complement C5 to Exacerbate Neuroinflammation and Neuronal Injury after Intracerebral Hemorrhage

doi: 10.1007/s10753-026-02456-z

Figure Lengend Snippet: ICH-Exos C5 promoted necroptosis and M1 polarization in hemin-induced microglia. ( A ) The heat map shows the differentially expressed proteins. ( B ) C5a level in brain tissue analyzed by IHC. * p < 0.05 vs. sham group. ( C ) C5a level in serum was detected by ELISA. * p < 0.05 vs. sham group. Under RIPA lysis conditions, the results of Western blot for C5a in ( D ) human RBC-Exos and ( E ) murine RBC-Exos after untreated EVs or after treatment with proteinase K (5 U/mL, 10 min) . # p < 0.05 vs. Sham-Exos/Normal-Exos group; @ p < 0.05 vs. ICH-Exos group. ( F ) C5a level in HMC3 cells was analyzed by Western blot and ELISA. ( G ) Cell viability was detected by CCK-8. ( H ) Cell apoptosis was detected by flow cytometry. ( I ) p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL, and MLKL levels were detected by Western blot. ( G ) The proportions of CD86 + M1 and CD163 + M2 macrophages were detected by flow cytometry. ( K ) iNOS, COX-2, CCL2, ARG1, CD206, and YM1 levels analyzed by qRT-PCR. ( L ) iNOS, COX-2, ARG1 and CD206 levels analyzed by Western blot. (M) ELISA was used to detect IL-1β, IL-6, and TNF-α levels. n = 3. # p < 0.05 vs. Normal-Exos group; & p < 0.05 vs. ICH-Exos group

Article Snippet: The mouse TNF-α (CSB-E04741m, CUSABIO, China), human TNF-α (KE00154, Proteintech), IL-1β (KE10003, KE00021, Proteintech), IL-6 (CSB-E04639m, CSB-E04638h, CUSABIO), and C5a (CSB-E08512h, CUSABIO) ELISA kits were operated according to the manufacturer’s instructions.

Techniques: Paraffin-embedded Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Lysis, Western Blot, CCK-8 Assay, Flow Cytometry, Quantitative RT-PCR

Journal: Cell reports

Article Title: Intracellular complement C5a/C5aR1 stabilizes β-catenin to promote colorectal tumorigenesis.

doi: 10.1016/j.celrep.2022.110851

Figure Lengend Snippet: Figure 6. CTSD cleaves C5 into C5a in CRC (A) RNA-seq analysis of the mRNA levels of various proteases, including Ctsd (red), in colonic tumors or the counterpart tissues of mice with or without AOM and DSS treatment, respectively. Data are represented as RPKM value for the 23 upregulated protease genes. (B) Kaplan-Meier analysis of the association between overall survival and the mRNA level of CTSD in tumor tissues of CRC patients (n = 364). Data were from The Cancer Genome Atlas (TCGA) database. (C) CTSD cleaved C5 to produce C5a in a dose-dependent manner, detected by Coomassie staining. (D and E) The mRNA (D) and protein (E) levels of CTSD in colons of AOM and DSS-treated or control mice detected by real-time PCR or immunoblotting assay, respectively. Data are represented as mean ± SEM; n R 4; **p < 0.01. (F) The local levels of mC5a in colonic tissue homogenates (CTHs) of mice with or without AOM and DSS treatment measured by ELISA. Data are represented as mean ± SEM; n R 4; *p < 0.05. (G) The effect of CTSD insufficiency on the intracellular C5a levels measured by ELISA. Data are represented as mean ± SD; n = 3; *p < 0.05; ***p < 0.001. (H) The effect of CTSD insufficiency on b-catenin protein levels. (I) A proposed model underlying the mechanism of intracellular C5a/C5aR1 signaling in colonic epithelial cells for regulating b-catenin stability via assembling a b-catenin stabilization complex of C5a/C5aR1/KCTD5/cullin3/Roc-1/b-catenin. See also Figure S7.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER PMX205 R&D Systems Cat#5196/1 Cycloheximide (CHX) CST Cat#2112 recombinant human cathepsin D protein R&D Systems Cat# 1014-AS-010 recombinant human KCTD5 protein (His Tag) Abcam Cat#ab137135 beta-Catenin Protein, Human, Recombinant (GST Tag) Sino Biological Cat#11279-H20B C5a Anaphylatoxin CompTech Cat#A144 C5 CompTech Cat#A120 Azoxymethane (AOM) Sigma-Aldrich Cat#A5486 Dextran Sulfate Sodium Salt (DSS 36,000–50,000 M.Wt) MP Biomedical CAS:9011-18-1 Critical commercial assays Human Complement Component C5a DuoSet ELISA kit R&D Systems Cat#DY2037 Mouse Complement Component C5a DuoSet ELISA kit R&D Systems Cat#DY2150 GTVision III immunohistochemical detection kit Gene Tech Cat#GK500705 KOD-Plus-Mutagenesis Kit TOYOBO Cat#SMK-101 Fast Silver Stain Kit Beyotime Cat#P0017S MinuteTM Cytoplasmic and Nuclear Fractionation kit Invent Biotechnologies Cat#SC-003 MinuteTM Endosome Isolation and Cell Fractionation Kit Invent Biotechnologies Cat#ED-028 Human Neutrophil Isolation Kit TBD Cat#LZS11131 TMB Substrate Set Biolegend Cat#421101 Stop Solution for TMB Substrate Biolegend Cat#423001 DAPI Fluoromount-G YEASEN Cat#36308ES11 Fixation Buffer Biolegend Cat#420801 Permeabilization Wash Buffer (10X) Biolegend Cat#421002 Protein A/G Magnetic Beads Bimake Cat#B23202 PrimeScriptTM RT Master Mix Takara Cat#RR036A TB Green Premix Ex TaqTM II Takara Cat#RR820A Deposited data RNA sequencing data NCBI GEO database GEO#GSE127946 4D-label free quantitative proteomics data This study See Tables S1 and S2 Protein profiling by LC-MS This study See Table S3 Experimental models: Cell lines HT-29 ATCC Cat#HTB-38 RKO ATCC Cat#CRL-2577 SW480 ATCC Cat#CCL-228 SW620 ATCC Cat#CCL-227 LoVo ATCC Cat#CCL-229 DLD-1 ATCC Cat#CCL-221 HCT 116 ATCC Cat#CCL-247 HCT-8 ATCC Cat#CCL-244 HCT-15 ATCC Cat#CCL-225 NCM460 Professor Guoxiang Cai N/A HCoEpiC Jennio-Bio N/A CT26.WT ATCC Cat#CRL-2638 HEK293FT Thermo Fisher Cat#R70007 HepG2 ATCC Cat#HB-8065 Experimental models: Organisms/strains C57BL/6 mice SLAC Laboratory Animal Co., Ltd N/A B10.D2-Hc0 mice The Jackson Laboratory Cat#000461 (Continued on next page) Cell Reports 39, 110851, May 31, 2022 e2

Techniques: RNA Sequencing, Staining, Control, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Intracellular complement C5a/C5aR1 stabilizes β-catenin to promote colorectal tumorigenesis.

doi: 10.1016/j.celrep.2022.110851

Figure Lengend Snippet: Figure 7. Correlation of C5a, C5aR1, b-catenin, and CTSD in human CRC specimens (A and B) Representative images of C5a, C5aR1, b-catenin, and CTSD staining in normal colon, adenoma, and adenocarcinoma tissues in clinical CRC spec- imens (A) or in paired CRC adenocarcinoma tissues and adjacent non-tumoral tissues (B). Scale bars, inner, 200 mm; outer, 50 mm. (C and D) Statistical analysis of the histological staining score (H score) in CRC specimens. Data are represented as min to max in (C) (related to A); n = 16 (normal), 35 (adenoma), or 9 (adenocarcinoma). Data of nine paired adenocarcinoma and adjacent tissues are shown in (D) (related to B); ns, not significant;***p < 0.001; ****p < 0.0001. (E and F) Scatter diagram analysis showing highly positive correlations of b-catenin with C5a or C5aR1 (E) or of CTSD with C5a or b-catenin (F).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER PMX205 R&D Systems Cat#5196/1 Cycloheximide (CHX) CST Cat#2112 recombinant human cathepsin D protein R&D Systems Cat# 1014-AS-010 recombinant human KCTD5 protein (His Tag) Abcam Cat#ab137135 beta-Catenin Protein, Human, Recombinant (GST Tag) Sino Biological Cat#11279-H20B C5a Anaphylatoxin CompTech Cat#A144 C5 CompTech Cat#A120 Azoxymethane (AOM) Sigma-Aldrich Cat#A5486 Dextran Sulfate Sodium Salt (DSS 36,000–50,000 M.Wt) MP Biomedical CAS:9011-18-1 Critical commercial assays Human Complement Component C5a DuoSet ELISA kit R&D Systems Cat#DY2037 Mouse Complement Component C5a DuoSet ELISA kit R&D Systems Cat#DY2150 GTVision III immunohistochemical detection kit Gene Tech Cat#GK500705 KOD-Plus-Mutagenesis Kit TOYOBO Cat#SMK-101 Fast Silver Stain Kit Beyotime Cat#P0017S MinuteTM Cytoplasmic and Nuclear Fractionation kit Invent Biotechnologies Cat#SC-003 MinuteTM Endosome Isolation and Cell Fractionation Kit Invent Biotechnologies Cat#ED-028 Human Neutrophil Isolation Kit TBD Cat#LZS11131 TMB Substrate Set Biolegend Cat#421101 Stop Solution for TMB Substrate Biolegend Cat#423001 DAPI Fluoromount-G YEASEN Cat#36308ES11 Fixation Buffer Biolegend Cat#420801 Permeabilization Wash Buffer (10X) Biolegend Cat#421002 Protein A/G Magnetic Beads Bimake Cat#B23202 PrimeScriptTM RT Master Mix Takara Cat#RR036A TB Green Premix Ex TaqTM II Takara Cat#RR820A Deposited data RNA sequencing data NCBI GEO database GEO#GSE127946 4D-label free quantitative proteomics data This study See Tables S1 and S2 Protein profiling by LC-MS This study See Table S3 Experimental models: Cell lines HT-29 ATCC Cat#HTB-38 RKO ATCC Cat#CRL-2577 SW480 ATCC Cat#CCL-228 SW620 ATCC Cat#CCL-227 LoVo ATCC Cat#CCL-229 DLD-1 ATCC Cat#CCL-221 HCT 116 ATCC Cat#CCL-247 HCT-8 ATCC Cat#CCL-244 HCT-15 ATCC Cat#CCL-225 NCM460 Professor Guoxiang Cai N/A HCoEpiC Jennio-Bio N/A CT26.WT ATCC Cat#CRL-2638 HEK293FT Thermo Fisher Cat#R70007 HepG2 ATCC Cat#HB-8065 Experimental models: Organisms/strains C57BL/6 mice SLAC Laboratory Animal Co., Ltd N/A B10.D2-Hc0 mice The Jackson Laboratory Cat#000461 (Continued on next page) Cell Reports 39, 110851, May 31, 2022 e2

Techniques: Staining