<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0609</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2026-09-16</periodOfReport>

    <issuer>
        <issuerCik>0001641281</issuerCik>
        <issuerName>Bolt Biotherapeutics, Inc.</issuerName>
        <issuerTradingSymbol>BOLT</issuerTradingSymbol>
        <issuerForeignTradingSymbol></issuerForeignTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001428666</rptOwnerCik>
            <rptOwnerName>Yonehiro Grant</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerNonUSAddressFlag>false</rptOwnerNonUSAddressFlag>
            <rptOwnerStreet1>C/O BOLT BIOTHERAPEUTICS, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>900 CHESAPEAKE DRIVE</rptOwnerStreet2>
            <rptOwnerCity>REDWOOD CITY</rptOwnerCity>
            <rptOwnerState>CA</rptOwnerState>
            <rptOwnerZipCode>94063</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>false</isDirector>
            <isOfficer>true</isOfficer>
            <isTenPercentOwner>false</isTenPercentOwner>
            <isOther>false</isOther>
            <officerTitle>Chief Operating Officer</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>false</aff10b5One>

    <nonDerivativeTable>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-09-16</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>1417</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.7740</value>
                    <footnoteId id="F1"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>29028</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-09-17</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>4909</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.3183</value>
                    <footnoteId id="F2"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>24119</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2026-09-18</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>6850</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>3.2364</value>
                    <footnoteId id="F3"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>17269</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
    </nonDerivativeTable>

    <footnotes>
        <footnote id="F1">Represents weighted average sales price. These shares were sold by the reporting person to cover tax withholding obligations in connection with the vesting of the restricted stock units granted on October 22, 2025. The shares were sold at prices ranging from $3.60 to $4.14. The Reporting Person will provide upon request, to the Securities and Exchange Commission (the &quot;SEC&quot;), the Issuer or security holder of the Issuer, full information regarding the number of shares sold at each separate price.</footnote>
        <footnote id="F2">Represents weighted average sales price. These shares were sold by the reporting person to cover tax withholding obligations in connection with the vesting of the restricted stock units granted on October 22, 2025. The shares were sold at prices ranging from $3.22 to $3.67. The Reporting Person will provide upon request, to the Securities and Exchange Commission (the &quot;SEC&quot;), the Issuer or security holder of the Issuer, full information regarding the number of shares sold at each separate price.</footnote>
        <footnote id="F3">Represents weighted average sales price. These shares were sold by the reporting person to cover tax withholding obligations in connection with the vesting of the restricted stock units granted on October 22, 2025. The shares were sold at prices ranging from $3.15 to $3.41. The Reporting Person will provide upon request, to the Securities and Exchange Commission (the &quot;SEC&quot;), the Issuer or security holder of the Issuer, full information regarding the number of shares sold at each separate price.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ William P. Quinn, Attorney-in-Fact</signatureName>
        <signatureDate>2026-09-21</signatureDate>
    </ownerSignature>
</ownershipDocument>
