KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-TLR2 | Abcam | Cat# ab1655; RRID: AB_302428 |
| Anti-TLR2 neutralizing | InvivoGen | Clone T2.5; Cat# mab-mtlr2; RRID: AB_763722 |
| Mouse Control IgG1 | InvivoGen | Cat# mabg1-ctrlm; RRID: AB_11203233 |
| Anti-C3 | MP-Biomedicals | Cat# 855444 |
| Anti-C3 | Abcam | Cat# ab11887; RRID: AB_298669 |
| Anti-C3d | Agilent | Cat# A006302; RRID: AB_578478 |
| Anti-CFB | Atlas Antibodies; Sigma | Cat# HPA001817; RRID: AB_1078779 |
| Anti-CFD | R&D systems | Cat# MAB5430; RRID: AB_10640506 |
| Rat IgG1 Isotype Control | R&D systems | Cat# MAB005; RRID: AB_357348 |
| MAC/ anti-C5b-9 for Mouse IHC | Biozol | Cat# FGI-10–1801 |
| MAC/ anti-C5b-9 for human IHC | Santa Cruz Biotechnology | clone aE11; Cat# sc58935; RRID: AB_1119839 |
| MAC/ anti-C5b-9 for WB | Santa Cruz Biotechnology | Clone 2A1; Cat# sc66190; RRID: AB_1119840 |
| Anti-CEP | Dr. Rowan (Rowan et al., 2017) | NA |
| ZO-1 | Thermo Fisher Scientific | Cat# 40–2200; RRID: AB_2533456 |
| IBA1 | Wako | Cat# 019–19741; RRID: AB_839504 |
| Biotinylated Goat Anti-Rabbit IgG | Vector Laboratories | Cat# BA-1000; RRID: AB_2313606 |
| Biotinylated Rabbit Anti-Goat IgG | Vector Laboratories | Cat# BA-5000; RRID: AB_2336126 |
| Anti-human CD16 | BioLegend | Clone 3G8; Cat# 302006; RRID: AB_314206 |
| Anti-human CD86 | Miltenyi Biotec | Clone FM95; Cat# 130–094-877; RRID: AB_10839702 |
| Anti-human CD45 | BioLegend | Clone 2D1; Cat# 368506; RRID: AB_2566358 |
| Anti-CD66b | BioLegend | Clone G10F5; Cat# 305116; RRID: AB_2566605 |
| Anti-human CD14 | Miltenyi | Clone Tük4; Cat# 130–098-058; RRID: AB_2660173 |
| Anti-human CD80 | Miltenyi | Clone 2D10; Cat# 130–099-710; RRID: AB_2659260 |
| Alexa Fluor 488 Phalloidin | Thermo Fisher Scientific | Cat# A12379; RRID: AB_2315147 |
| Alexa Fluor 647 Donkey Anti-Mouse | Abcam | Cat# AB150107 |
| Alexa Fluor 488 Goat Anti-Rabbit | Thermo Fisher Scientific | Cat #A11034; RRID: AB_2576217 |
| Alexa Fluor 488 Goat anti-Mouse | Thermo Fisher Scientific | Cat #A11001; RRID: AB_2534069 |
| Biological Samples | ||
| Human donor eye tissue | The Iowa Lions Eye Bank | https://iowalionseyebank.org/ |
| Peripheral Blood Mononuclear Cells (PBMCs) | Isolated from healthy volunteers | NA |
| Primary Microglia | Isolated as per Fernando et al., 2016 | NA |
| Normal Human Serum | Sigma | Cat# H4522–20ml |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Mouse M-CSF | Miltenyl Biotec | Cat# 130–094-129 |
| Mouse Recombinant GM-CSF | Stem Cell Technologies | Cat# 78017.1 |
| Pam3Cys4 | Invivogen | Cat# tlrl-pms |
| CEP-HSA | Prof. Salomon (Gu et al., 2003) | |
| Mal (TIRAP) Inhibitor Peptide | Calbiochem | Cat# 613570–1MG |
| Sodium Iodate | Sigma | Cat # S4007–100G |
| Hoechst | Sigma | Cat #B2261–25MG |
| Collagen IV | Sigma | Cat# C5533–5M |
| Critical Commercial Assays | ||
| In SITU Cell Death Detection Kit, TMR red | Roche | Cat# 12156792910 |
| MCP-1 ELISA | Tebubio | Cat# 900-K31 |
| MAC ELISA | Abbexa | Cat# abx350654 |
| Isolate II RNA Extraction Kit | Bioline | Cat# BIO-52073 |
| MMLV Reverse Transcriptase | Promega | Cat# M1705 |
| SensiFast SYBER Green | Bioline | Cat# BIO-92020 |
| Pierce LDH Viability Assay | Thermo Fisher Scientific | Cat# 13464269 |
| LIVE/DEAD Aqua | Thermo Fisher Scientific | Cat# L34957 |
| Mouse Vectastain ABC HRP Kit | Vector Laboratories | Cat#PK-4002; RRID: AB_2336811 |
| ImmPACT DAB Substrate | Vector Laboratories | Cat#SK-4105; RRID: AB_2336520 |
| Vectastain Elite ABC HRP kit | Vector Laboratories | Cat#PK-6200; RRID: AB_2336826 |
| Vector VIP Peroxidase (HRP) Substrate Kit | Vector Laboratories | Cat# SK-4600; RRID: AB_2336848 |
| Experimental Models: Cell Lines | ||
| ARPE19 | ATCC | CRL-2302 |
| Immortalized BMDMS | Prof. Golenbock, UMASS, USA | N/A |
| hfRPE | Dr. A. Maminishkis (Maminishkis et al., 2006) | N/A |
| THP-1 | ATCC | TIB-202 |
| HEK293-TLR2 | Dr. K. Fitzgerald, UMASS, USA | N/A |
| Experimental Models: Organisms/Strains | ||
| C57Bl6 | Jackson labs | |
| Tlr2–/– | Jackson labs | JAX stock #004650 |
| DBA/2J | Jackson labs | JAX stock #000671 |
| Oligonucleotides | ||
| Primers for SYBER qPCR, | This paper | N/A |
| Recombinant DNA | ||
| C3 luciferase (T1 del) | Addgene | |
| Software and Algorithms | ||
| GraphPad Prism | GraphPad Software | https://www.graphpad.com |
| ImageJ | https://imagej.nih.gov/ij/ | |
| FlowJo | Tree star | https://www.flowjo.com |